5-HT4 receptor agonist compounds

ABSTRACT

The invention provides novel quinolinone-carboxamide 5-HT 4  receptor agonist compounds. The invention also provides pharmaceutical compositions comprising such compounds, methods of using such compounds to treat diseases associated with 5-HT 4  receptor activity, and processes and intermediates useful for preparing such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser.No. 11/266,881, filed Nov. 4, 2005, now U.S. Pat. No. 7,399,862 whichapplication claims the benefit of U.S. Provisional Application No.60/625,185, filed on Nov. 5, 2004, the entire disclosures of which areincorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention is directed to quinolinone-carboxamide compounds which areuseful as 5-HT₄ receptor agonists. The invention is also directed topharmaceutical compositions comprising such compounds, methods of usingsuch compounds for treating medical conditions mediated by 5-HT₄receptor activity, and processes and intermediates useful for preparingsuch compounds.

2. State of the Art

Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter that iswidely distributed throughout the body, both in the central nervoussystem and in peripheral systems. At least seven subtypes of serotoninreceptors have been identified and the interaction of serotonin withthese different receptors is linked to a wide variety of physiologicalfunctions. There has been, therefore, substantial interest in developingtherapeutic agents that target specific 5-HT receptor subtypes.

In particular, characterization of 5-HT₄ receptors and identification ofpharmaceutical agents that interact with them has been the focus ofsignificant recent activity. (See, for example, the review by Langloisand Fischmeister, J. Med. Chem. 2003, 46, 319-344.) 5-HT₄ receptoragonists are useful for the treatment of disorders of reduced motilityof the gastrointestinal tract. Such disorders include irritable bowelsyndrome (IBS), chronic constipation, functional dyspepsia, delayedgastric emptying, gastroesophageal reflux disease (GERD), gastroparesis,post-operative ilcus, intestinal pseudo-obstruction, and drug-induceddelayed transit. In addition, it has been suggested that some 5-HT₄receptor agonist compounds may be used in the treatment of centralnervous system disorders including cognitive disorders, behavioraldisorders, mood disorders, and disorders of control of autonomicfunction.

Despite the broad utility of pharmaceutical agents modulating 5-HT₄receptor activity, few 5-HT₄ receptor agonist compounds are in clinicaluse at present.

Accordingly, there is a need for new 5-HT₄ receptor agonists thatachieve their desired effects with minimal side effects. Preferredagents may possess, among other properties, improved selectivity,potency, pharmacokinetic properties, and/or duration of action.

SUMMARY OF THE INVENTION

The invention provides novel compounds that possess 5-HT₄ receptoragonist activity. Among other properties, compounds of the inventionhave been found to be potent and selective 5-HT₄ receptor agonists.

Accordingly, the invention provides a compound of formula (I):

or a pharmaceutically-acceptable salt or solvate or stereoisomerthereof,

wherein:

R¹ is hydrogen, halo, hydroxy, C₁₋₄alkyl, or C₁₋₄alkoxy;

R² is C₃₋₄alkyl or C₃₋₆cycloalkyl;

R³ is hydrogen or C₁₋₃alkyl;

R⁴ is —S(O)₂—C₁₋₃alkyl, —C(O)O—C₁₋₃alkyl or —C(O)—C₁₋₃alkyl;

n is an integer of 0, 1, 2, or 3;

Y is selected from:

-   -   (a) a moiety of formula (a)

wherein:

Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸,—N(R⁸)C(O)OR^(d), —N(R⁸)C(O)NR^(e)R^(f), —N(R⁸)SO₂NR^(g)R^(h),—C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —C(O)OR^(m), —OR⁸, —SR^(n), cyano,hydroxy-substituted C₁₋₄alkyl, hydroxy-substituted C₁₋₃alkoxy, —CF₃,pyridinyl, thiomorpholinyl, thiazolidinyl, imidazolyl, indolyl,tetrahydrofuranyl, pyrrolidinyl and piperidinyl, wherein pyrrolinyl isoptionally substituted with oxo and piperidinyl is optionallysubstituted with 1 to 3 halo;

R⁵ is selected from hydrogen and C₁₋₄alkyl, wherein C₁₋₄alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,halo, and C₁₋₃alkoxy;

m is an integer of 0, 1, 2, 3, 4, or 5;

R⁶ and R⁷ are independently selected from hydrogen, hydroxy, halo andC₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2substituents selected from hydroxy, C₁₋₃alkoxy, and cyano;

R⁸ is independently hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,C₁₋₃alkoxy, and cyano;

R^(a) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionallysubstituted with —SO₂R^(b), C₃₋₆cycloalkyl or with from 1 to 3 halo;

R^(c) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionallysubstituted with 1 to 2 substituents selected from hydroxy andC₃₋₆cycloalkyl, or with from 1 to 3 halo;

and R^(b), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i), R^(j), R^(k),R^(l), R^(m), and R^(n), are independently hydrogen or C₁₋₄alkyl;

or R⁵ and R⁶, R⁵ and R⁸, or R⁶ and R⁸, taken together form aC₂₋₅alkylene, wherein the C₂₋₅alkylene is optionally substituted with 1to 2 substituents selected from hydroxy, halo and C₁₋₄alkyl, whereinC₁₋₄alkyl is optionally substituted with 1 to 2 substituents selectedfrom hydroxy, C₁₋₃alkoxy, and cyano;

or R⁸ and R^(a) taken together form a C₂₋₅alkylene;

provided that when m is 1, Z forms a carbon-carbon bond with the carbonatom bearing the substituents R⁶ and R⁷; and when m is 0, Z is selectedfrom —S(O₂)R⁸, —C(O)NR^(i)R^(j), —C(O)OR^(m), and —CF₃;

(b) —OR⁹, wherein R⁹ is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,C₁₋₃alkoxy, and cyano;

(c) —SR¹⁰, wherein R¹⁰ is hydrogen or C₁₋₄alkyl; and

(d) a heterocycle or heteroaryl selected from pyridinyl,thiomorpholinyl, thiazolidinyl, imidazolyl, indolyl, tetrahydrofuranyl,pyrrolidinyl and piperidinyl, wherein pyrrolinyl is optionallysubstituted with oxo and piperidinyl is optionally substituted with 1 to3 halo;

provided that when n is 0, Y is selected from (d), wherein theheterocycle or heteroaryl is attached via a carbon atom of theheterocycle or heteroaryl ring.

The invention also provides a pharmaceutical composition comprising acompound of the invention and a pharmaceutically-acceptable carrier.

The invention also provides a method of treating a disease or conditionassociated with 5-HT₄ receptor activity, e.g. a disorder of reducedmotility of the gastrointestinal tract, the method comprisingadministering to the mammal, a therapeutically effective amount of acompound of the invention.

Further, the invention provides a method of treating a disease orcondition associated with 5-HT₄ receptor activity in a mammal, themethod comprising administering to the mammal, a therapeuticallyeffective amount of a pharmaceutical composition of the invention.

The invention provides a method of treating irritable bowel syndrome,the method comprising administering to the mammal, a therapeuticallyeffective amount of a pharmaceutical composition of the invention.

The compounds of the invention can also be used as research tools, i.e.to study biological systems or samples, or for studying the activity ofother chemical compounds. Accordingly, in another of its method aspects,the invention provides a method of using a compound of formula (I), or apharmaceutically acceptable salt or solvate or stereoisomer thereof, asa research tool for studying a biological system or sample or fordiscovering new 5-HT₄ receptor agonists, the method comprisingcontacting a biological system or sample with a compound of theinvention and determining the effects caused by the compound on thebiological system or sample.

In separate and distinct aspects, the invention also provides syntheticprocesses and novel intermediates described herein, which are useful forpreparing compounds of the invention.

The invention also provides a compound of the invention as describedherein for use in medical therapy, as well as the use of a compound ofthe invention in the manufacture of a formulation or medicament fortreating a disease or condition associated with 5-HT₄ receptor activity,e.g. a disorder of reduced motility of the gastrointestinal tract, in amammal.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides novel quinolinone-carboxamide 5-HT₄ receptoragonists of formula (I), or pharmaceutically-acceptable salts orsolvates or stereoisomers thereof. The following exemplary and preferredvalues for radicals, substituents, and ranges, are for illustrationonly; they do not exclude other defined values or other values withindefined ranges for the radicals and substituents unless otherwiseindicated.

In a specific aspect of the invention, R¹ is hydrogen, halo, C₁₋₄alkyl,or C₁₋₄alkoxy.

In other specific aspects, R¹ is hydrogen, halo, or C₁₋₃alkyl; or R¹ isfluoro; or R¹ is bromo.

In another specific aspect, R¹ is hydrogen.

In a specific aspect, R² is C₃₋₄alkyl. Representative R² groups includen-propyl, isopropyl, n-butyl, sec-butyl, and tert-butyl.

In another specific aspect, R² is isopropyl.

In yet another specific aspect, R² is cyclobutyl or cyclopentyl.

In a specific aspect, R³ is hydrogen.

In another specific aspect, R³ is methyl.

In a specific aspect, R⁴ is —S(O)₂—C₁₋₃alkyl. In another specificaspect, R⁴ is —S(O)₂CH₃.

In yet another specific aspect, R⁴ is —C(O)O—C₁₋₃alkyl. RepresentativeR⁴ groups include —C(O)OCH₃, —C(O)OCH₂CH₃, —C(O)OCH(CH₃)₂, and—C(O)OCH₂CH₂CH₃.

In another specific aspect, R⁴ is —C(O)OCH₃.

In yet another specific aspect, R⁴ is —C(O)—C₁₋₃alkyl. Representative R⁴groups include —C(O)CH₃, —C(O)CH₂CH₃, —C(O)CH(CH₃)₂, and —C(O)CH₂CH₂CH₃.

In another specific aspect, R⁴ is —C(O)CH₃.

In yet another specific aspect, R⁴ is selected from —S(O)₂CH₃,—C(O)OCH₃, and —C(O)CH₃.

In a specific aspect, n is an integer of 0, 1, or 2. In another specificaspect, n is an integer of 1, 2, or 3. In other specific aspects, n is 1or 2; or n is 1.

In a specific aspect, Y is selected from:

(a) a moiety of formula (a), wherein Z is selected from —N(R⁸)SO₂R^(a),—N(R⁸)C(O)R^(c), —S(O₂)R⁸, —N(R⁸)C(O)NR^(e)R^(f), —C(O)NR^(i)R^(j),—OC(O)NR^(k)R^(l), —OR⁸, and cyano;

(b) —O—C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to2 substituents selected from hydroxy;

(c) —S—C₁₋₄alkyl; and

(d) pyridinyl, imidazolyl, indolyl, and tetrahydrofuranyl, whereinpyrrolidinyl is optionally substituted with oxo.

In another specific aspect of the invention, Y is a moiety of formula(a).

In another specific aspect of the invention, Y is —OR⁹.

In yet another specific aspect, Y is —O—C₁₋₃alkyl, wherein C₁₋₃alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,C₁₋₃alkoxy, and cyano. In other specific aspects of the invention, Y is—SR¹⁰; or Y is —S—C₁₋₃alkyl.

In yet another aspect of the invention, Y is a heterocycle or heteroarylselected from pyridinyl, thiomorpholinyl, thiazolidinyl, imidazolyl,indolyl, tetrahydrofuranyl, pyrrolidinyl and piperidinyl, whereinpyrrolinyl is optionally substituted with oxo and piperidinyl isoptionally substituted with 1 to 3 halo. In another specific aspect, Yis selected from pyridinyl, imidazolyl, indolyl, and tetrahydrofuranyl,wherein pyrrolidinyl is optionally substituted with oxo.

In specific aspects, Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c),—S(O₂)R⁸, —N(R⁸)C(O)OR^(d), —N(R⁸)C(O)NR^(e)R^(f), —N(R⁸)SO₂NR^(g)R^(h),—C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —C(O)OR^(m), —OR⁸, —SR^(n), andcyano; or Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸,—N(R⁸)C(O)NR^(e)R^(f), —C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —OR⁸, andcyano.

In another specific aspect, Z is selected from —N(R⁸)SO₂R^(a),—N(R⁸)C(O)R^(c), —S(O₂)R⁸, and —N(R⁸)C(O)NR^(e)R^(f).

In still another specific aspect, Z is —N(R⁸)SO₂R^(a).

In a specific aspect, R⁵ is C₁₋₄alkyl wherein C₁₋₄ alkyl is optionallysubstituted with 1 to 2 substituents selected from hydroxy, halo, andC₁₋₃alkoxy. In one aspect, R⁵ is methyl. In another specific aspect, R⁵is hydrogen.

In a specific aspect, m is an integer of 0, 1, 2, 3 or 4. In anotherspecific aspect, m is an integer of 1, 2, 3 or 4. In other specificaspects, m is 0, 1, 2 or 3; m is 1, 2 or 3; m is 2, 3 or 4; m is 2 or 3;or m is 2.

In a specific aspect, R⁶ and R⁷ are independently selected from hydrogenand C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2substituents selected from hydroxy, C₁₋₃alkoxy, and cyano. In otherspecific aspects, R⁶ and R⁷ are independently hydrogen and methyl; or R⁶and R⁷ are hydrogen.

In a specific aspect, R⁸ is selected from hydrogen and C₁₋₄alkyl, suchas hydrogen and methyl.

In a specific aspect, R^(a) and R^(c) are independently selected fromhydrogen and C₁₋₄alkyl.

In a specific aspect, R^(b), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i),R^(j), R^(k), R^(l), R^(m), and R^(n) are independently selected fromhydrogen and methyl.

In another specific aspect of the invention, R⁵ and R⁶, or R⁵ and R⁸taken together form an optionally substituted C₂₋₅alkylene.

In another specific aspect, R⁵ and R⁶, or R⁵ and R⁸ taken together formethylene, or propylene.

In a specific aspect, R⁶ and R⁸ taken together form an optionallysubstituted C₂₋₅alkylene.

In another specific aspect, R⁶ and R⁸ taken together form ethylene, orpropylene.

In a specific aspect, R⁸ and R^(a) taken together form a C₂₋₅alkylene.

In a specific aspect, R⁹ is an optionally substituted C₁₋₃alkyl.

In a specific aspect, R¹⁰ is C₁₋₃alkyl.

In a specific aspect, the invention provides a compound of formula (I)wherein R¹ is hydrogen, R² is C₃₋₄alkyl, and R³ is hydrogen.

In another specific aspect, the invention provides a compound of formula(I) wherein R¹ is hydrogen or halo; R² is isopropyl or C₄₋₅cycloalkyl;R³ is hydrogen or methyl, and R⁴, n and Y are as defined herein.

In another specific aspect, the invention provides a compound of formula(I) wherein n is 1, and when Y is a moiety of formula (a), m is 1, 2 or3.

In yet another specific aspect, the invention provides a compound offormula (I) wherein:

-   -   R¹ is hydrogen;    -   R² is C₃₋₄alkyl;    -   R³ is hydrogen;    -   Y is selected from:    -   (a) a moiety of formula (a):

-   -   wherein Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c),        —S(O₂)R⁸, —N(R⁸)C(O)NR^(e)R^(f), —C(O)NR^(i)R^(j),        —OC(O)NR^(k)R^(l), —OR⁸, and cyano;    -   (b) —O—C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted        with 1 to 2 substituents selected from hydroxy;    -   (c) —S—C₁₋₄alkyl; and    -   (d) pyridinyl, imidazolyl, indolyl, and tetrahydrofuranyl,        wherein pyrrolidinyl is optionally substituted with oxo;

and R⁴, n, m, R⁵, R⁶, R⁷, R⁸, R^(a), R^(c), R^(e), R^(f), R^(i), R^(j),R^(k), and R^(l) are as defined herein.

In yet another specific aspect, the invention further provides acompound of formula (I) which is a compound of formula (II):

wherein

R¹ is hydrogen, halo, or C₁₋₃alkyl;

R² is C₃₋₄alkyl;

R³ is hydrogen or methyl,

R⁴ is —S(O)₂—C₁₋₃alkyl, —C(O)O—C₁₋₃alkyl, or —C(O)—C₁₋₃alkyl;

n is an integer of 1 or 2;

R⁵ is selected from hydrogen and C₁₋₄alkyl, wherein C₁₋₄ alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,halo, and C₁₋₃alkoxy;

m is an integer of 1, 2, 3, 4, or 5;

R⁶ and R⁷ are independently selected from hydrogen and C₁₋₄alkyl,wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituentsselected from hydroxy, C₁₋₃alkoxy, and cyano;

Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸,—N(R⁸)C(O)OR^(d), —N(R⁸)C(O)NR^(e)R^(f), —N(R⁸)SO₂NR^(g)R^(h),—C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —C(O)OR^(m), —OR⁸, —SR^(n), andcyano;

R⁸ is independently hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl isoptionally substituted with 1 to 2 substituents selected from hydroxy,C₁₋₃alkoxy, and cyano;

R^(a) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionallysubstituted with —SO₂R^(b), C₃₋₆cycloalkyl or with from 1 to 3 halo;

R^(c) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionallysubstituted with 1 to 2 substituents selected from hydroxy andC₃₋₆cycloalkyl, or with from 1 to 3 halo;

and R^(b), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i), R^(j), R^(k),R^(l), R^(m), and R^(n), are independently hydrogen or C₁₋₄alkyl;

or R⁵ and R⁶, R⁵ and R⁸, or R⁶ and R⁸, taken together form aC₂₋₅alkylene, wherein the C₂₋₅alkylene is optionally substituted with 1to 2 substituents selected from hydroxy, halo and C₁₋₄alkyl, whereinC₁₋₄alkyl is optionally substituted with 1 to 2 substituents selectedfrom hydroxy, C₁₋₃alkoxy, and cyano;

or R⁸ and R^(a) taken together form a C₂₋₅alkylene.

In still other aspects, the invention provides the compounds listed inTables 1 to 6 herein, i.e., compounds of formulae (II-a), (II-b),(II-c), (II-d), (II-e), and (II-f).

The chemical naming conventions used herein are illustrated for thecompound of Example 2:

which is designated 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide,according to the AutoNom software, provided by MDL Information Systems,GmbH (Frankfirt, Germany). The designation (1S,3R,5R) describes therelative orientation of the bonds associated with the bicyclic ringsystem that are depicted as solid and dashed wedges. The compound isalternatively denoted asN-[(3-endo)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl]-1-(1-methylethyl)-2-oxo-1,2-dihydro-3-quinolinecarboxamide.In all of the compounds of the invention listed by name below, thequinolinone carboxamide is endo to the azabicyclooctane group.

Particular mention may be made of the following compounds:

-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-hydroxy-3-[(3-imidazol-1-yl-propyl)methanesulfonylamino]propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-aza-bicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxy-propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-{2-[3-(methanesulfonylmethylamino)piperidin-1-yl]ethyl}-carbamic    acid methyl ester;-   [2-(3-acetylaminopiperidin-1-yl)ethyl]-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamic    acid methyl ester;-   [2-(3-acetylaminopyrrolidin-1-yl)ethyl]-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamic    acid methyl ester;-   (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(3-dimethylaminosulfonyl-aminopyrrolidin-1-yl)-ethyl]-carbamic    acid methyl ester;-   (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)-amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(3-dimethylaminosulfonyl    methylaminopyrrolidin-1-yl)-ethyl]-carbamic acid methyl ester;-   {2-[3-(1,1-dioxo-1λ⁶-isothiazolidin-2-yl)pyrrolidin-1-yl]ethyl}-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-aza-bicyclo[3.2.1]oct-8-yl}propyl)-carbamic    acid methyl ester;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    [(1S,3R,5R)-8-(3-{acetyl-[2-(3-carbamoylpiperidin-1-yl)ethyl]amino}-2-hydroxypropyl)-8-azabicyclo[3.2.1]oct-3-yl]-amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(acetyl-{2-[4-(methanesulfonylaminomethyl)piperidin-1-yl]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide;-   (1-{2-[acetyl-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)amino]-ethyl}pyrrolidin-3-yl)-carbamic    acid methyl ester;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(acetyl-{2-[3-(dimethylaminosulfonylmethylamino)pyrrolidin-1-yl]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(acetyl-{2-[3-(trimethylureido)pyrrolidin-1-yl]ethyl}amino)-2-hydroxy-propyl]-8-azabicyclo-[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(acetyl-{2-[(1-dimethylsulfamoylpyrrolidin-3-yl)methylamino]ethyl}amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[acetyl-(2-{[2-(methanesulfonylmethylamino)ethyl]methylamino}-ethyl)amino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(acetyl-{2-[(2-methanesulfonyl amino    ethyl)methylamino]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide;    and-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[acetyl-(2-{[2-(1,1-dioxo-1λ⁶-isothiazolidin-2-yl)ethyl]methylamino}ethyl)amino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide.

As exemplified by particular compounds listed above, the compounds ofthe invention may contain one or more chiral centers. Accordingly, theinvention includes racemic mixtures, pure stereoisomers, andstereoisomer-enriched mixtures of such isomers, unless otherwiseindicated. When a particular stereoisomer is shown, it will beunderstood by those skilled in the art, that minor amounts of otherstereoisomers may be present in the compositions of the invention unlessotherwise indicated, provided that any utility of the composition as awhole is not eliminated by the presence of such other isomers.

DEFINITIONS

When describing the compounds, compositions and methods of theinvention, the following terms have the following meanings, unlessotherwise indicated.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched or combinations thereof. Examples ofparticular values for a C₁₋₄alkyl group include, by way of example,methyl, ethyl, n-propyl (n-Pr), isopropyl (i-Pr), n-butyl (n-Bu),sec-butyl, isobutyl, and tert-butyl.

The term “alkylene” means a divalent saturated hydrocarbon group whichmay be linear or branched or combinations thereof. Examples ofparticular values for a C₂₋₅alkylene include ethylene, propylene,isopropylene, butylene, and pentylene, and the like.

The term “alkoxy” means the monovalent group —O-alkyl, where alkyl isdefined as above. Representative alkoxy groups include, by way ofexample, methoxy, ethoxy, propoxy, butoxy, and the like.

The term “cycloalkyl” means a monovalent saturated carbocyclic groupwhich may be monocyclic or multicyclic. Unless otherwise defined, suchcycloalkyl groups typically contain from 3 to 10 carbon atoms.Representative C₃₋₆cycloalkyl groups include, by way of example,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term “compound” means a compound that was synthetically prepared orprepared in any other way, such as by metabolism.

The term “halo” means a fluoro, chloro, bromo or iodo.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need of treatment.

The term “treatment” as used herein means the treatment of a disease,disorder, or medical condition in a patient, such as a mammal(particularly a human) which includes:

-   -   (a) preventing the disease, disorder, or medical condition from        occurring, i.e., prophylactic treatment of a patient;    -   (b) ameliorating the disease, disorder, or medical condition,        i.e., eliminating or causing regression of the disease,        disorder, or medical condition in a patient;    -   (c) suppressing the disease, disorder, or medical condition,        i.e., slowing or arresting the development of the disease,        disorder, or medical condition in a patient; or    -   (d) alleviating the symptoms of the disease, disorder, or        medical condition in a patient.

The term “pharmaceutically-acceptable salt” means a salt prepared froman acid or base which is acceptable for administration to a patient,such as a mammal. Such salts can be derived frompharmaceutically-acceptable inorganic or organic acids and frompharmaceutically-acceptable bases. Typically,pharmaceutically-acceptable salts of compounds of the present inventionare prepared from acids.

Salts derived from pharmaceutically-acceptable acids include, but arenot limited to, acetic, benzenesulfonic, benzoic, camphorsulfonic,citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic,hydrochloric, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pantothenic, phosphoric, succinic, sulfuric, tartaric,p-toluenesulfonic, xinafoic (1-hydroxy-2-naphthoic acid),napthalene-1,5-disulfonic acid and the like.

The term “solvate” means a complex or aggregate formed by one or moremolecules of a solute, i.e. a compound of the invention or apharmaceutically-acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include by way of example, water, methanol, ethanol,isopropanol, acetic acid, and the like. When the solvent is water, thesolvate formed is a hydrate.

It will be appreciated that the term “or a pharmaceutically-acceptablesalt or solvate of stereoisomer thereof” is intended to include allpermutations of salts, solvates and stereoisomers, such as a solvate ofa pharmaceutically-acceptable salt of a stereoisomer of a compound offormula (I).

The term “amino-protecting group” means a protecting group suitable forpreventing undesired reactions at an amino nitrogen. Representativeamino-protecting groups include, but are not limited to, formyl; acylgroups, for example alkanoyl groups, such as acetyl; alkoxycarbonylgroups, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl groups,such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc);arylmethyl groups, such as benzyl (Bn), trityl (Tr), and1,1-di-(4′-methoxyphenyl)methyl; silyl groups, such as trimethylsilyl(TMS) and tert-butyldimethylsilyl (TBDMS); and the like.

The term “hydroxy protecting group” means a protecting group suitablefor preventing undesired reactions of a hydroxyl group. The term“hydroxyl-protecting group” means a protecting group suitable forpreventing undesirable reactions at a hydroxyl group. Representativehydroxyl-protecting groups include, but are not limited to, silyl groupsincluding tri(1-6C)alkylsilyl groups, such as trimethylsilyl (TMS),triethylsilyl (TES), tert-butyldimethylsilyl (TBS) and the like; esters(acyl groups) including (1-6C)alkanoyl groups, such as formyl, acetyland the like; arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl(PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM) and thelike. Additionally, two hydroxyl groups can also be protected as analkylidene group, such as prop-2-ylidine, formed, for example, byreaction with a ketone, such as acetone.

General Synthetic Procedures

Compounds of the invention can be prepared from readily availablestarting materials using the following general methods and procedures.Although a particular aspect of the present invention is illustrated inthe schemes below, those skilled in the art will recognize that allaspects of the present invention can be prepared using the methodsdescribed herein or by using other methods, reagents and startingmaterials known to those skilled in the art. It will also be appreciatedthat where typical or preferred process conditions (i.e., reactiontemperatures, times, mole ratios of reactants, solvents, pressures,etc.) are given, other process conditions can also be used unlessotherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group, as well assuitable conditions for protection and deprotection, are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and G. M. Wuts, ProtectingGroups in Organic Synthesis, Third Edition, Wiley, New York, 1999, andreferences cited therein.

The substituents and variables shown in the following schemes have thedefinitions provided herein unless otherwise indicated.

In one method of synthesis, compounds of formula (I) are prepared asillustrated in Scheme A:

In Scheme A, a compound of formula (III) wherein R¹, R², R³, n and Y areas defined herein, is reacted with a compound of formula (IV) wherein L¹is a leaving group, such as halo, for example, chloro, or ethoxy, orL¹-R⁴ is a carboxylic acid, i.e., L¹ represents a hydroxy group.

Optimal reaction conditions for the reaction of Scheme A may varydepending on the chemical properties of the reagent L¹-R⁴, as is wellknown to those skilled in the art.

For example, when L¹ is a halo leaving group, such as chloro, thereaction is typically conducted by contacting a compound of formula(III) with between about 1 and about 4 equivalents of a compound offormula (IV) in an inert diluent, such as dichloromethane, in thepresence of an excess of a base, for example, between about 3 and about6 equivalents of base, such as N,N-diisopropylethylamine or1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Suitable inert diluents alsoinclude N,N-dimethylformamide (DMF), trichloromethane,1,1,2,2-tetrachloroethane, tetrahydrofuran, and the like. The reactionis typically conducted at a temperature in the range of about −100° C.to about 30° C. for about a quarter hour to about 2 hours, or until thereaction is substantially complete. Exemplary reagents L¹-R⁴ includemethanesulfonylchloride and acetylchloride, and the like.

When the reagent L¹-R⁴ is a carboxylic acid, Scheme A represents anamide coupling reaction which is typically conducted by contacting acompound of formula (III) with between about 1 and about 4 equivalentsof a carboxylic acid in an inert diluent, for example,N,N-dimethylformamide, in the presence of a coupling agent such asbenzotriazol-1-yloxytripyrrolidino-phosphonium hexafluorophosphate(PyBOP). The reaction is typically conducted at ambient temperature, forabout a quarter hour to about 2 hours, or until the reaction issubstantially complete. Suitable alternative coupling agents include 1,3dicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodimide (EDC), and PyBOP combinedwith 1-hydroxy-7-azabenzotriazole (HOAt).

The amide coupling of a compound of formula (III) with a carboxylic acidalternatively can be performed by converting the carboxylic acid to anactivated ester, such as an N-hydroxy succinimide (NHS) ester or ap-nitrophenyl ester, or an acid imidazole, which is then reacted with acompound of formula (III).

When the reagent L¹-R⁴ is a liquid, for example ethyl formate, thereaction can be performed by dissolving a compound of formula (III) in alarge excess of the reagent L¹-R⁴, and heating to a temperature ofbetween about 50° C. and about 100° C. for about 12 to about 24 hours.

The product of formula (I) is isolated and purified by conventionalprocedures. For example, the product can be concentrated to drynessunder reduced pressure, taken up in an aqueous weak acid solution andpurified by HPLC chromatography.

Similarly, compounds of formula (I) can also be prepared by N-alkylatinga compound of the form of formula (I) in which R² is hydrogen, which canbe prepared by the schemes described herein, except substituting theappropriate reagents. The N-alkylation reaction is typically conductedby contacting a compound of the form of formula (I) in which R² ishydrogen, with between about 1 and about 4 equivalents of a compound ofthe formula L²-R² in which L² is a leaving group such as halo (chloro,iodo or bromo), or a sulfonic ester group, such as mesylate, tosylate,brosylate, nosylate and the like; and R² is C₃₋₄alkyl or C₃₋₆cycloalkyl.This reaction is typically conducted in a polar aprotic solvent such asdimethylformamide in the presence of between about 2 and about 4equivalents of strong base, such as potassium tert-butoxide. Typically,the reaction is conducted at a temperature of between about 60° C. andabout 100° C. for between about 6 and about 24 hours, or until thereaction is substantially complete.

In yet another alternative, compounds of formula (I) in which R¹ isother than hydrogen are prepared by conventional processes fromcompounds of formula (I) in which R¹ is hydrogen.

In another method of synthesis, compounds of formula (II) can beprepared as illustrated in Scheme B:

by reductive amination of an aldehyde of formula (V) or a hydrate of analdehyde (V). The reductive amination is typically conducted bycontacting a compound of formula (V) with between about 1 and about 6equivalents of an amine compound of formula (VI) in the presence of areducing agent. Suitable reducing agents, for example, include hydrogenin the presence of a Group VIII metal catalyst, such as palladium oncharcoal, or a borohydride, such as sodium triacetoxyborohydride, sodiumcyanoborohydride, lithium cyanoborohydride, and the like. Convenientsolvents include halogenated hydrocarbons, such as dichloromethane(DCM), and alcohols, such as methanol. Typically, the reaction isconducted at a temperature of between about 0° C. and about 40° C. forbetween about 0.5 hour and about 4 hours, or until the reaction issubstantially complete.

Optimal reaction conditions for the reaction of Scheme B may varydepending on the chemical properties of a compound of formula (VI), asis well known to those skilled in the art. Compounds of formula (VI) areavailable from commercial sources or can be prepared from readilyavailable starting materials, as discussed in the Examples containedherein.

A compound of formula (III) can be prepared as illustrated in Scheme C:

wherein P² is an amino-protecting group; and L³ and L⁴ are leavinggroups.

A negatively-charged counterion is also present associated with thepositively-charged intermediate compound (5) or (5′).

A substituted quinolinone carboxylic acid (1) can be readily prepared byprocedures similar to those reported in the literature in Suzuki et al,Heterocycles, 2000, 53, 2471-2485 and described in the examples below.

A protected aminotropane (2) or aminoazabicyclooctane can be preparedfrom readily available starting materials. For example, when theprotecting group P² is Boc, the protected tropane can be prepared bycontacting 2,5-dimethoxy tetrahydrofuran with between about 1 and 2equivalents, preferably about 1.5 equivalents of benzyl amine and aslight excess, for example about 1.1 equivalents, of1,3-acetonedicarboxylic acid in an acidic aqueous solution in thepresence of a buffering agent such as sodium hydrogen phosphate. Thereaction mixture is heated to between about 60° C. and about 100° C. toensure decarboxylation of any carboxylated intermediates in the product,8-benzyl-8-azabicyclo[3.2.1]octan-3-one, commonly N-benzyltropanone. Theproduct is typically reacted with a slight excess of di-tert-butyldicarbonate, for example, about 1.1 equivalents, under a hydrogenatmosphere in the presence of a transition metal catalyst to provide aBoc protected intermediate, 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester. The reaction is typically conducted at ambienttemperature for about 12 to about 72 hours. Finally, the ester iscontacted with a large excess, for example between about 10 to about 40equivalents, of ammonium formate in an inert diluent, such as methanol,in the presence of a transition metal catalyst to provide anaminotropane in the endo configuration, typically, more than 99% in theendo configuration. The product can be purified by conventionalprocedures, such as by alkaline extraction.

Intermediate compound (3) can be prepared by coupling a substitutedquinolinone carboxylic acid (1) with a protected aminotropane (2) underconditions similar to those described in Scheme A for amide bondformation. The protecting group P² can be removed by standard proceduresto provide an intermediate compound (3). For example when the protectinggroup is Boc, typically removal is by treatment with an acid, such astrifluoroacetic acid, providing the acid salt of the intermediate. Theacid salt of intermediate compound (3) can be converted to the freebase, if desired, by conventional treatment with base. The protectinggroup Cbz, for another example, is conveniently removed byhydrogenolysis over a suitable metal catalyst, such as palladium oncarbon.

It will be understood that in the reaction described above, and in otherprocesses described herein using intermediate compound (3), intermediatecompound (3) can be supplied in the form of the free base or in a saltform, with appropriate adjustment of reaction conditions, as necessary,as known to those skilled in the art.

An azetidine intermediate (5) can be prepared by reacting intermediatecompound (3) with an oxirane compound, wherein L³ represents a leavinggroup such as bromo, chloro, or iodo, to form an azetidine salt offormula (5). The oxirane compound can be, for example,2-bromomethyloxirane (commonly, epibromohydrin). This reaction istypically conducted by contacting intermediate compound (3) with betweenabout 2 and about 4 equivalents of 2-bromomethyloxirane in a polardiluent, such as ethanol. The reaction is typically conducted at ambienttemperature for between about 24 and about 48 hours or until thereaction is substantially complete.

An azetidine intermediate of formula (5′), in which R³ is C₁₋₃alkyl, canbe prepared by contacting intermediate (5) with from slightly less thanone equivalent to about one equivalent of a compound of formula L⁴-R³,where R³ is C₁₋₃alkyl, and L⁴ is a leaving group, such as halo (chloro,bromo, or iodo) or the like; in an inert diluent in the presence ofbetween about 1 and about 3 equivalents of a strong base, such aspotassium tert-butoxide or sodium hydride. The reaction is typicallyconducted at ambient temperature for between about a quarter hour to anhour, or until the reaction is substantially complete. Suitable inertdiluents include dichloromethane, trichloromethane,1,1,2,2-tetrachloroethane, and the like.

A compound of formula (III) can be prepared by reacting an azetidinecompound of formula (5) or (5′) with an amine compound of formula (6) inthe presence of a base to provide a compound of formula (III).Typically, the azetidine intermediate is dissolved in an inert diluent,such as methanol, in the presence of a base, and contacted with betweenabout 1 and about 8 equivalents, such as between about 1 and 3equivalents, of the amine. The reaction is typically conducted at atemperature of between about 0° C. and about 100° C. for between about12 and about 24 hours or until the reaction is substantially complete.

A compound of formula (V) can be prepared as illustrated in Scheme D:

In Scheme D, an azetidine intermediate (5) or (5′), wherein R³ ishydrogen or C₁₋₃alkyl respectively, is reacted with an amino acetal (7).The acetal intermediate (8) product is then reacted with a compound offormula (IV), L¹-R⁴, wherein L¹ is a leaving group, to provide anaminoacetal intermediate (9). The aminoacetal intermediate (9) is thenhydrolyzed to form an aldehyde compound of formula (V) or the hydrate ofaldehyde (V). The process of Scheme D is described further in Examples3-4.

A solution of an azetidine intermediate in an inert diluent, such asethanol, is mixed with an amino acetal (7), such as aminoacetaldehydedimethylacetal, in the presence of a base. Suitable bases include DIPEA,DBU and the like. The mixture is typically refluxed and stirred for fromabout 6 hours to about 20 hours, then concentrated under reducedpressure to provide acetal intermediate (8).

Typically, a solution of intermediate (8) in an inert diluent, such asdichloromethane, in the presence of a base, such as DIPEA, is reactedwith between about 1 and about 3 equivalents of compound (IV), L¹-R⁴,(where L¹ is a leaving group such as halo, and R⁴ is —C(O)O—C₁₋₃alkyl or—C(O)—C₁₋₃alkyl). The reaction is typically conducted at between about−20° C. and about 20° C. for between about 0.5 hours to about 12 hours.The stirred reaction is allowed to warm to room temperature. Thereaction mixture is concentrated and purified to provide intermediate(9).

The acetal intermediate (9) is typically contacted with an acid, such asaqueous hydrochloric acid, and stirred for between about 30 minutes andabout 2 hours. Then the excess reagent is removed. The reaction mixtureis diluted with aqueous aectonitrile then lyophilized to provide acompound of formula (V).

Alternatively, compounds of formula (I) can be prepared as illustratedin Scheme E shown below:

by reacting a compound of formula (VII) wherein L⁵ is a leaving group,such as halo, with a compound of formula (VIII).

Optimal reaction conditions for the reaction of Scheme E may varydepending on the chemical properties of the reagent H—Y, as is wellknown to those skilled in the art.

For example, when L⁵ is a halo leaving group, such as chloro, and H—Y isan N-linked heterocycle, the reaction is typically conducted bycontacting a compound of formula (VII) with between about 1 and about 4equivalents of reagent H—Y in an inert diluent, such as methanol,ethanol, DMF, N-methylpyrrolinone (NMP), and the like, in the presenceof a base, such as N,N-diisopropylethylamine or1,8-diazabicyclo[5.4.0]-undec-7-ene (DBU). Suitable inert diluentsinclude nitriles, such as acetonitrile, N,N-dimethylformamide,1,1,2,2-tetrachloroethane, tetrahydrofuran, and the like. The reactionis typically conducted at a temperature in the range of about 40° C. toabout 120° C. for about 2 hours to about 24 hours, or until the reactionis substantially complete.

A compound of formula (VII) can be prepared by reducing a compound offormula (V) to an alcohol in the presence of a reducing agent, such ashydrogen and a metal catalyst, or a borohydride, such as sodiumborohydride, then reacting the alcohol product with a reagent, such asphosphorus tribromide or thionyl chloride, to provide a compound offormula (VII).

Alternatively, when R⁴ is —S(O)₂—C₁₋₃alkyl, a compound of formula (I)can be prepared according to Scheme F:

wherein L⁶ is a leaving group, such as halo, and the asterisk denotes achiral center. The process utilizing a compound of formula (IX) isuseful for preparing compounds of formula (I) in which thestereochemistry at the carbon marked by the asterisk is specifically (R)or (S) as well as for preparing non-chiral compounds.

Typically, a compound of formula (IX) is contacted with between about 1and about 6 equivalents of compound (X) in a polar diluent, such asmethanol, ethanol, DMF, NMP, or the like, in the presence of 2 to 3equivalents of a base, such as N,N-diisopropylethylamine, potassiumcarbonate, sodium hydroxide, and the like. The reaction is typicallyconducted at a temperature of between about 0° C. and about 120° C. forbetween about 12 and about 24 hours, or until the reaction issubstantially complete to provide a compound of formula (I).

In an alternative method of synthesis, a compound of formula (I) can beprepared as illustrated in Scheme G:

wherein L⁷ is a leaving group, such as halo, and the asterisk denotes achiral center.

Alternatively, a compound of formula (I) wherein R³ is hydrogen, and theasterisk denotes a chiral center, can be prepared as illustrated inScheme H:

Schemes G and H are useful for preparing compounds of formulae (I) inwhich the stereochemistry at the center marked by the asterisk isspecifically (R) or (S) as well as for preparing non-chiral compounds.

Typically, a compound of formula (3) is contacted with between about 1and about 4 equivalents of a compound of formula (XI) in a polardiluent, such as methanol, N,N-dimethylformamide (DMF), or the like, inthe presence of more than one equivalent of a base, such asN,N-diisopropylethylamine or the like. Alternatively, a compound offormula (3) is contacted with between about 1 and about 4 equivalents ofa compound of formula (XII) in a polar diluent, such as methanol,N,N-dimethylformamide (DMF), or the like. The reaction is typicallyconducted at a temperature of between about 25° C. and about 100° C. forbetween about 2 and about 24 hours, or until the reaction issubstantially complete to provide a compound of formula (I).

In yet another alternative process, a compound of formula (I) can beprepared according to Scheme J:

The reaction of Scheme J is typically conducted under amide couplingconditions known in the art. Typically, this reaction is conducted byconverting a carboxylic acid (i.e., a compound of formula (1)) to anacid chloride by contacting compound (1) with at least one equivalent,preferably between about 1 and about 2 equivalents of an activatingagent such as thionyl chloride or oxalyl chloride in an aromaticdiluent, such as toluene, benzene, xylene, or the like. The reaction istypically conducted at a temperature ranging from about 80° C. to about120° C. for about 15 minutes to about 4 hours, or until the reaction issubstantially complete.

The acid chloride solution is typically added to a biphasic mixture ofabout 1 equivalent of the aminotropane (a compound of formula (XIII)) toform a compound of formula (I). The biphasic mixture of a compound offormula (XIII) is generally prepared by dissolving a compound of formula(XIII) in an aromatic diluent, such as toluene, and adding an aqueoussolution containing an excess of base, such as sodium hydroxide orpotassium hydroxide, preferably about 2 to about 10 equivalents of base.

Alternatively, the amide coupling of a compound of formula (XIII) withthe carboxylic acid compound of formula (1) can be performed in thepresence of a coupling agent such as 1,3 dicyclohexylcarbodiimide (DCC),1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (EDC), orbenzotriazol-1-yloxytripyrrolidino-phosphonium hexafluorophosphate(PyBop), optionally combined with 1-hydroxy-7-azabenzotriazole (HOAt),as described above in Scheme A for the amide coupling of a compound offormula (III) with a carboxylic acid (L¹-R⁴). In yet anotheralternative, the amide coupling of Scheme J can be performed byconverting a compound of formula (1) to an activated ester, as describedin Scheme A herein.

In yet another alternative process, when R⁴ is —C(O)O—C₁₋₃alkyl or—C(O)—C₁₋₃alkyl a compound of formula (I) can be prepared according toScheme K:

by reacting an azetidine compound of formula (5) or (5′) with a compoundof formula (XIV) in the presence of a strong base, such as sodiumhydride. Typically, the azetidine intermediate is dissolved in a polardiluent, such as DMF, in the presence of a strong base, and contactedwith between about 1 and about 8 equivalents, such as between about 1and 3 equivalents, of the protected amine. The reaction is typicallyconducted at a temperature of between about 0° C. and about 100° C. forbetween about 12 and about 24 hours or until the reaction issubstantially complete.

Reagents L¹-R⁴, L²-R², L⁴-R³, H—Y, compounds of formulae (IX), (X),(XI), (XII), (XIII), and (XIV), and other intermediates are readilyprepared by standard procedures from common starting materials or areavailable commercially. Further details regarding specific reactionconditions and other procedures for preparing representative compoundsof the invention or intermediates thereto are described in the examplesbelow.

Accordingly, in a method aspect, the invention provides a process forpreparing a compound of formula (I), or a pharmaceutically-acceptablesalt or solvate or stereoisomer thereof, the process comprising:

(a) reacting a compound of formula (III):

or a salt or stereoisomer or protected derivative thereof;

with a compound of formula (IV):L¹-R⁴  (IV)

wherein L¹ is a leaving group; or

(b) reacting a compound of formula (VII)

wherein L⁵ is a leaving group; or a salt or stereoisomer or protectedderivative thereof;

with a compound of formula (VIII):H—Y  (VIII)

to provide a compound of formula (I) or a pharmaceutically-acceptablesalt or solvate or stereoisomer thereof.

The invention further provides a process for preparing a compound offormula (II), or a pharmaceutically-acceptable salt or solvate orstereoisomer thereof, the process comprising:

reacting a compound of formula (V):

or a salt, hydrate, stereoisomer or protected derivative thereof;

with a compound of formula (VI):

in the presence of a reducing agent to provide a compound of formula(II) or a pharmaceutically-acceptable salt or solvate or stereoisomerthereof.

In other embodiments, this invention is directed to the other processesdescribed herein; and to the products prepared by any of the processesdescribed herein.

The invention further provides a compound of formula (III), or a salt orstereoisomer or protected derivative thereof. In a specific aspect, theinvention provides a compound of formula (III), or a salt orstereoisomer or protected derivative thereof, wherein Y is a moiety offormula (a) and R¹, R², R³, and n are as defined herein.

Pharmaceutical Compositions

The quinolinone-carboxamide compounds of the invention are typicallyadministered to a patient in the form of a pharmaceutical composition.Such pharmaceutical compositions may be administered to the patient byany acceptable route of administration including, but not limited to,oral, rectal, vaginal, nasal, inhaled, topical (including transdermal)and parenteral modes of administration.

Accordingly, in one of its compositions aspects, the invention isdirected to a pharmaceutical composition comprising apharmaceutically-acceptable carrier or excipient and a therapeuticallyeffective amount of a compound of formula (I) or a pharmaceuticallyacceptable salt thereof. Optionally, such pharmaceutical compositionsmay contain other therapeutic and/or formulating agents if desired.

The pharmaceutical compositions of the invention typically contain atherapeutically effective amount of a compound of the present inventionor a pharmaceutically-acceptable salt thereof. Typically, suchpharmaceutical compositions will contain from about 0.1 to about 95% byweight of the active agent; preferably, from about 5 to about 70% byweight; and more preferably from about 10 to about 60% by weight of theactive agent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of the invention. The choice of a particular carrier orexcipient, or combinations of carriers or excipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable pharmaceutical composition for a particular mode ofadministration is well within the scope of those skilled in thepharmaceutical arts. Additionally, the ingredients for such compositionsare commercially available from, for example, Sigma, P.O. Box 14508, St.Louis, Mo. 63178. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of pharmacy, 20^(th) Edition, Lippincott Williams & White,Baltimore, Md. (2000); and H. C. Ansel et al., Pharmaceutical DosageForms and Drug Delivery Systems, 7^(th) Edition, Lippincott Williams &White, Baltimore, Md. (1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following: (1)sugars, such as lactose, glucose and sucrose; (2) starches, such as cornstarch and potato starch; (3) cellulose, such as microcrystallinecellulose, and its derivatives, such as sodium carboxymethyl cellulose,ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5)malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter andsuppository waxes; (9) oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10)glycols, such as propylene glycol; (11) polyols, such as glycerin,sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyloleate and ethyl laurate; (13) agar; (14) buffering agents, such asmagnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19)ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxiccompatible substances employed in pharmaceutical compositions.

The pharmaceutical compositions of the invention are typically preparedby thoroughly and intimately mixing or blending a compound of theinvention with a pharmaceutically-acceptable carrier and one or moreoptional ingredients. If necessary or desired, the resulting uniformlyblended mixture can then be shaped or loaded into tablets, capsules,pills and the like using conventional procedures and equipment.

The pharmaceutical compositions of the invention are preferably packagedin a unit dosage form. The term “unit dosage form” means a physicallydiscrete unit suitable for dosing a patient, i.e., each unit containinga predetermined quantity of active agent calculated to produce thedesired therapeutic effect either alone or in combination with one ormore additional units. For example, such unit dosage forms may becapsules, tablets, pills, and the like.

In a preferred embodiment, the pharmaceutical compositions of theinvention are suitable for oral administration. Suitable pharmaceuticalcompositions for oral administration may be in the form of capsules,tablets, pills, lozenges, cachets, dragees, powders, granules; or as asolution or a suspension in an aqueous or non-aqueous liquid; or as anoil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup;and the like; each containing a predetermined amount of a compound ofthe present invention as an active ingredient.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the pharmaceutical compositionsof the invention will typically comprise a compound of the presentinvention as the active ingredient and one or morepharmaceutically-acceptable carriers, such as sodium citrate ordicalcium phosphate. Optionally or alternatively, such solid dosageforms may also comprise: (1) fillers or extenders, such as starches,microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as carboxymethylcellulose, alginates,gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants,such as glycerol; (4) disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and/or sodium carbonate; (5) solution retarding agents, such asparaffin; (6) absorption accelerators, such as quaternary ammoniumcompounds; (7) wetting agents, such as cetyl alcohol and/or glycerolmonostearate; (8) absorbents, such as kaolin and/or bentonite clay; (9)lubricants, such as talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and/or mixtures thereof;(10) coloring agents; and (11) buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants can also be presentin the pharmaceutical compositions of the invention. Examples ofpharmaceutically-acceptable antioxidants include: (1) water-solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodiumbisulfate, sodium metabisulfate sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal-chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like. Coating agents fortablets, capsules, pills and like, include those used for entericcoatings, such as cellulose acetate phthalate (CAP), polyvinyl acetatephthalate (PVAP), hydroxypropyl methylcellulose phthalate, methacrylicacid-methacrylic acid ester copolymers, cellulose acetate trimellitate(CAT), carboxymethyl ethyl cellulose (CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), and the like.

If desired, the pharmaceutical compositions of the present invention mayalso be formulated to provide slow or controlled release of the activeingredient using, by way of example, hydroxypropyl methyl cellulose invarying proportions; or other polymer matrices, liposomes and/ormicrospheres.

In addition, the pharmaceutical compositions of the present inventionmay optionally contain opacifying agents and may be formulated so thatthey release the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.Examples of embedding compositions which can be used include polymericsubstances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically-acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Such liquid dosage formstypically comprise the active ingredient and an inert diluent, such as,for example, water or other solvents, solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (such as, for example, cottonseed, groundnut,corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Suspensions, in addition to the active ingredient, maycontain suspending agents such as, for example, ethoxylated isostearylalcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

Alternatively, the pharmaceutical compositions of the invention areformulated for administration by inhalation. Suitable pharmaceuticalcompositions for administration by inhalation will typically be in theform of an aerosol or a powder. Such compositions are generallyadministered using well-known delivery devices, such as a metered-doseinhaler, a dry powder inhaler, a nebulizer or a similar delivery device.

When administered by inhalation using a pressurized container, thepharmaceutical compositions of the invention will typically comprise theactive ingredient and a suitable propellant, such asdichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas.

Additionally, the pharmaceutical composition may be in the form of acapsule or cartridge (made, for example, from gelatin) comprising acompound of the invention and a powder suitable for use in a powderinhaler. Suitable powder bases include, by way of example, lactose orstarch.

The compounds of the invention can also be administered transdermallyusing known transdermal delivery systems and excipients. For example, acompound of the invention can be admixed with permeation enhancers, suchas propylene glycol, polyethylene glycol monolaurate,azacycloalkan-2-ones and the like, and incorporated into a patch orsimilar delivery system. Additional excipients including gelling agents,emulsifiers and buffers, may be used in such transdermal compositions ifdesired.

The following formulations illustrate representative pharmaceuticalcompositions of the present invention:

FORMULATION EXAMPLE A

Hard gelatin capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 50 mg Lactose (spray-dried)200 mg  Magnesium stearate 10 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then loaded into a hard gelatin capsule (260 mg of        composition per capsule).

FORMULATION EXAMPLE B

Hard gelatin capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 20 mg Starch 89 mgMicrocrystalline cellulose 89 mg Magnesium stearate  2 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then passed through a No. 45 mesh U.S. sieve and loaded into        a hard gelatin capsule (200 mg of composition per capsule).

FORMULATION EXAMPLE C

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 10 mg Polyoxyethylenesorbitan monooleate 50 mg Starch powder 250 mg 

-   -   Representative Procedure: The ingredients are thoroughly blended        and then loaded into a gelatin capsule (310 mg of composition        per capsule).

FORMULATION EXAMPLE D

Tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 5 mg Starch 50 mg Microcrystalline cellulose 35 mg  Polyvinylpyrrolidone (10 wt. % inwater) 4 mg Sodium carboxymethyl starch 4.5 mg   Magnesium stearate 0.5mg   Talc 1 mg

-   -   Representative Procedure: The active ingredient, starch and        cellulose are passed through a No. 45 mesh U.S. sieve and mixed        thoroughly. The solution of polyvinylpyrrolidone is mixed with        the resulting powders, and this mixture is then passed through a        No. 14 mesh U.S. sieve. The granules so produced are dried at        50-60° C. and passed through a No. 18 mesh U.S. sieve. The        sodium carboxymethyl starch, magnesium stearate and talc        (previously passed through a No. 60 mesh U.S. sieve) are then        added to the granules. After mixing, the mixture is compressed        on a tablet machine to afford a tablet weighing 100 mg.

FORMULATION EXAMPLE E

Tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 25 mg Microcrystallinecellulose 400 mg  Silicon dioxide fumed 10 mg Stearic acid  5 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then compressed to form tablets (440 mg of composition per        tablet).

FORMULATION EXAMPLE F

Single-scored tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 15 mg Cornstarch 50 mgCroscarmellose sodium 25 mg Lactose 120 mg  Magnesium stearate  5 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and compressed to form a single-scored tablet (215 mg of        compositions per tablet).

FORMULATION EXAMPLE G

A suspension for oral administration is prepared as follows:

Ingredients Amount Compound of the invention 0.1 g Fumaric acid 0.5 gSodium chloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 gGranulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum k(Vanderbilt Co.) 1.0 g Flavoring 0.035 mL Colorings 0.5 mg Distilledwater q.s. to 100 mL

-   -   Representative Procedure: The ingredients are mixed to form a        suspension containing 10 mg of active ingredient per 10 mL of        suspension.

FORMULATION EXAMPLE H

A dry powder for administration by inhalation is prepared as follows:

Ingredients Amount Compound of the invention 1.0 mg Lactose  25 mg

-   -   Representative Procedure: The active ingredient is micronized        and then blended with lactose. This blended mixture is then        loaded into a gelatin inhalation cartridge. The contents of the        cartridge are administered using a powder inhaler.

FORMULATION EXAMPLE I

A dry powder for administration by inhalation in a metered dose inhaleris prepared as follows:

-   -   Representative Procedure: A suspension containing 5 wt. % of a        compound of the invention and 0.1 wt. % lecithin is prepared by        dispersing 10 g of active compound as micronized particles with        mean size less than 10 μm in a solution formed from 0.2 g of        lecithin dissolved in 200 mL of demineralized water. The        suspension is spray dried and the resulting material is        micronized to particles having a mean diameter less than 1.5 μm.        The particles are loaded into cartridges with pressurized        1,1,1,2-tetrafluoroethane.

FORMULATION EXAMPLE J

An injectable formulation is prepared as follows:

Ingredients Amount Compound of the invention 0.2 g Sodium acetate buffersolution (0.4 M)  40 mL HCl (0.5 N) or NaOH (0.5 N) q.s. to pH 4 Water(distilled, sterile) q.s. to 20 mL

-   -   Representative Procedure: The above ingredients are blended and        the pH is adjusted to 4±0.5 using 0.5 N HCl or 0.5 N NaOH.

FORMULATION EXAMPLE K

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the Invention 4.05 mg Microcrystallinecellulose (Avicel PH 103) 259.2 mg  Magnesium stearate 0.75 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then loaded into a gelatin capsule (Size #1, White, Opaque)        (264 mg of composition per capsule).

FORMULATION EXAMPLE L

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the Invention   8.2 mg Microcrystallinecellulose (Avicel PH 103) 139.05 mg Magnesium stearate  0.75 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then loaded into a gelatin capsule (Size #1, White, Opaque)        (148 mg of composition per capsule).

It will be understood that any form of the compounds of the invention,(i.e. free base, pharmaceutical salt, or solvate) that is suitable forthe particular mode of administration, can be used in the pharmaceuticalcompositions discussed above.

Utility

The quinolinone-carboxamide compounds of the invention are 5-HT₄receptor agonists and therefore are expected to be useful as therapeuticagents for treating medical conditions mediated by 5-HT₄ receptors orassociated with 5-HT₄ receptor activity, i.e. medical conditions whichare ameliorated by treatment with a 5-HT₄ receptor agonist. Such medicalconditions include, but are not limited to, irritable bowel syndrome(IBS), chronic constipation, functional dyspepsia, delayed gastricemptying, gastroesophageal reflux disease (GERD), gastroparesis,diabetic and idiopathic gastropathy, post-operative ileus, intestinalpseudo-obstruction, and drug-induced delayed transit. In addition, ithas been suggested that some 5-HT₄ receptor agonist compounds may beused in the treatment of central nervous system disorders includingcognitive disorders, behavioral disorders, mood disorders, and disordersof control of autonomic function.

In particular, the compounds of the invention are expected to be usefulfor treating disorders of the GI tract caused by reduced motility inmammals, including humans. Such GI motility disorders include, by way ofillustration, chronic constipation, irritable bowel syndrome, diabeticand idiopathetic gastroparesis, and functional dyspepsia.

When used to treat disorders of reduced motility of the GI tract orother conditions mediated by 5-HT₄ receptors, the compounds of theinvention will typically be administered orally in a single daily doseor in multiple doses per day, although other forms of administration maybe used. The amount of active agent administered per dose or the totalamount administered per day will typically be determined by a physician,in the light of the relevant circumstances, including the condition tobe treated, the chosen route of administration, the actual compoundadministered and its relative activity, the age, weight, and response ofthe individual patient, the severity of the patient's symptoms, and thelike.

Suitable doses for treating disorders of reduced motility of the GItract or other disorders mediated by 5-HT₄ receptors will range fromabout 0.0007 to about 20 mg/kg/day of active agent, preferably fromabout 0.0007 to about 1 mg/kg/day. For an average 70 kg human, thiswould amount to from about 0.05 to about 70 mg per day of active agent.

In one aspect of the invention, the compounds of the invention are usedto treat chronic constipation. When used to treat chronic constipation,the compounds of the invention will typically be administered orally ina single daily dose or in multiple doses per day. Preferably, the dosefor treating chronic constipation will range from about 0.05 to about 70mg per day.

In another aspect of the invention, the compounds of the invention areused to treat irritable bowel syndrome. When used to treatconstipation-predominant irritable bowel syndrome, the compounds of theinvention will typically be administered orally in a single daily doseor in multiple doses per day. Preferably, the dose for treatingconstipation-predominant irritable bowel syndrome will range from about0.05 to about 70 mg per day.

In another aspect of the invention, the compounds of the invention areused to treat diabetic gastroparesis. When used to treat diabeticgastroparesis, the compounds of the invention will typically beadministered orally in a single daily dose or in multiple doses per day.Preferably, the dose for treating diabetic gastroparesis will range fromabout 0.05 to about 70 mg per day.

In yet another aspect of the invention, the compounds of the inventionare used to treat functional dyspepsia. When used to treat functionaldyspepsia, the compounds of the invention will typically be administeredorally in a single daily dose or in multiple doses per day. Preferably,the dose for treating functional dyspepsia will range from about 0.05 toabout 70 mg per day.

The invention also provides a method of treating a mammal having adisease or condition associated with 5-HT₄ receptor activity, the methodcomprising administering to the mammal a therapeutically effectiveamount of a compound of the invention or of a pharmaceutical compositioncomprising a compound of the invention.

Since compounds of the invention are 5-HT₄ receptor agonists, suchcompounds are also useful as research tools for investigating orstudying biological systems or samples having 5-HT₄ receptors, or fordiscovering new 5-HT₄ receptor agonists. Moreover, since compounds ofthe invention exhibit binding selectivity for 5-HT₄ receptors ascompared with binding to receptors of other 5-HT subtypes, particularly5-HT₃ receptors, such compounds are particularly useful for studying theeffects of selective agonism of 5-HT₄ receptors in a biological systemor sample. Any suitable biological system or sample having 5-HT₄receptors may be employed in such studies which may be conducted eitherin vitro or in vivo. Representative biological systems or samplessuitable for such studies include, but are not limited to, cells,cellular extracts, plasma membranes, tissue samples, mammals (such asmice, rats, guinea pigs, rabbits, dogs, pigs, etc.) and the like.

In this aspect of the invention, a biological system or samplecomprising a 5-HT₄ receptor is contacted with a 5-HT₄ receptor-agonizingamount of a compound of the invention. The effects of agonizing the5-HT₄ receptor are then determined using conventional procedures andequipment, such as radioligand binding assays and functional assays.Such functional assays include ligand-mediated changes in intracellularcyclic adenosine monophosphate (cAMP), ligand-mediated changes inactivity of the enzyme adenylyl cyclase (which synthesizes cAMP),ligand-mediated changes in incorporation of analogs of guanosinetriphosphate (GTP), such as [³⁵S]GTPγS (guanosine5′-O-(γ-thio)triphosphate) or GTP-Eu, into isolated membranes viareceptor catalyzed exchange of GTP analogs for GDP analogs,ligand-mediated changes in free intracellular calcium ions (measured,for example, with a fluorescence-linked imaging plate reader or FLIPR®from Molecular Devices, Inc.), and measurement of mitogen activatedprotein kinase (MAPK) activation. A compound of the invention mayagonize or increase the activation of 5-HT₄ receptors in any of thefunctional assays listed above, or assays of a similar nature. A 5-HT₄receptor-agonizing amount of a compound of the invention will typicallyrange from about 1 nanomolar to about 1000 nanomolar.

Additionally, the compounds of the invention can be used as researchtools for discovering new 5-HT₄ receptor agonists. In this embodiment,5-HT₄ receptor binding or functional data for a test compound or a groupof test compounds is compared to the 5-HT₄ receptor binding orfunctional data for a compound of the invention to identify testcompounds that have superior binding or functional activity, if any.This aspect of the invention includes, as separate embodiments, both thegeneration of comparison data (using the appropriate assays) and theanalysis of the test data to identify test compounds of interest.

Among other properties, compounds of the invention have been found to bepotent agonists of the 5-HT₄ receptor and to exhibit substantialselectivity for the 5-HT₄ receptor subtype over the 5-HT₃ receptorsubtype in radioligand binding assays. In addition, representativecompounds have been shown not to exhibit an unacceptable level ofinhibition of the potassium ion current in an in vitro voltage-clampmodel using isolated whole cells expressing the hERG cardiac potassiumchannel. The voltage-clamp assay is an accepted pre-clinical method ofassessing the potential for pharmaceutical agents to change the patternof cardiac repolarization, specifically to cause, so-called QTprolongation, which has been associated with cardiac arrhythmia. (Caveroet al., Opinion on Pharmacotherapy, 2000, 1, 947-73, Fermini et al.,Nature Reviews Drug Discovery, 2003, 2, 439-447) Accordingly,pharmaceutical compositions comprising compounds of the invention areexpected to have an acceptable cardiac profile.

These properties, as well as the utility of the compounds of theinvention, can be demonstrated using various in vitro and in vivo assayswell-known to those skilled in the art. Representative assays aredescribed in further detail in the following examples.

EXAMPLES

The following synthetic and biological examples are offered toillustrate the invention, and are not to be construed in any way aslimiting the scope of the invention. In the examples below, thefollowing abbreviations have the following meanings unless otherwiseindicated. Abbreviations not defined below have their generally acceptedmeanings.

-   -   Boc=tert-butoxycarbonyl    -   (Boc)₂O=di-tert-butyl dicarbonate    -   DCM=dichloromethane    -   DMF=N,N-dimethylformamide    -   DMSO=dimethyl sulfoxide    -   EtOAc=ethyl acetate    -   mCPBA=m-chloroperbenzoic acid    -   MeCN=acetonitrile    -   MTBE=tert-butyl methyl ether    -   PyBOP=benzotriazol-1-yloxytripyrrolidino-phosphonium        hexafluorophosphate    -   R_(f)=retention factor    -   RT=room temperature    -   TFA=trifluoroacetic acid    -   THF=tetrahydrofuran

Reagents (including some secondary amines) and solvents were purchasedfrom commercial suppliers (Aldrich, Fluka, Sigma, etc.), and usedwithout further purification. Reactions were run under nitrogenatmosphere, unless noted otherwise. Progress of reaction mixtures wasmonitored by thin layer chromatography (TLC), analytical highperformance liquid chromatography (anal. HPLC), and mass spectrometry,the details of which are given below and separately in specific examplesof reactions. Reaction mixtures were worked up as described specificallyin each reaction; commonly they were purified by extraction and otherpurification methods such as temperature-, and solvent-dependentcrystallization, and precipitation. In addition, reaction mixtures wereroutinely purified by preparative HPLC: a general protocol is describedbelow. Characterization of reaction products was routinely carried outby mass and ¹H-NMR spectrometry. For NMR measurement, samples weredissolved in deuterated solvent (CD₃OD, CDCl₃, or DMSO-d₆), and ¹H-NMRspectra were acquired with a Varian Gemini 2000 instrument (300 MHz)under standard observation conditions. Mass spectrometric identificationof compounds was performed by an electrospray ionization method (ESMS)with a Perkin Elmer instrument (PE SCIEX API 150 EX).

General Protocol for Analytical HPLC

Crude compounds were dissolved in 50% MeCN/H₂O (with 0.1% TFA) at0.5-1.0 mg/mL concentration, and analyzed using the followingconditions:

Column: Zorbax Bonus-RP (3.5 μm of particle size, 2.1 × 50 mm) Flowrate: 0.5 mL/min Mobile A = 90% MeCN/10% H₂O/0.1% TFA Phases: B = 98%H₂O/2% MeCN/0.1% TFA Gradient: 10% A/90% B (0-0.5 min); 10% A/90% Bto50% A/50% B (linear, 0.5-5 min) Detector 214, 254, and 280 nm.wavelength:Alternative conditions, when used, are indicated explicitly.General Protocol for Preparative HPLC Purification

Crude compounds were dissolved in 50% acetic acid in water at 50-100mg/mL concentration, filtered, and fractionated using the followingprocedure:

Column: YMC Pack-Pro C18 (50 a × 20 mm; ID = 5 μm) Flow rate: 40 mL/minMobile Phases: A = 90% MeCN/10% H₂O/0.1% TFA B = 98% H₂O/2% MeCN/0.1%TFA Gradient: 10% A/90% B to 50% A/50% B over 30 min (linear) Detector214 nm. wavelength:Preparation of Secondary Amines (such as Compounds of Formula (VIII),H—Y)

Preparation of Various Secondary Amines Used as Intermediates in theSynthesis of a compound of formula (I) are described below.

Thiomorpholine-1,1-dioxide was prepared from thiomorpholine byprotection of the secondary amine to N-Boc thiomorpholine ((Boc)₂O,MeOH), oxidation to sulfone (mCPBA, CH₂Cl₂, 0° C.), and deprotection ofthe N-Boc group to provide the free amine (CF₃CO₂H, CH₂Cl₂). (m/z):[M+H]⁺ calcd for C₄H₉NO₂S, 136.04; found, 135.9.

The N-sulfonyl derivatives of piperazine were prepared from N-Bocpiperazine by reacting with respective sulfonyl chloride (iPr₂NEt,CH₂Cl₂, 0° C.), and deprotecting the N-Boc group (CF₃CO₂H, CH₂Cl₂).1-Methanesulfonyl-piperazine: ¹H-NMR (CDCl₃; neutral): δ (ppm) 3.1 (t,4H), 2.9 (t, 4H), 2.7 (s, 3H).1-(Methylsulfonyl)methanesulfonyl-piperazine: ¹H-NMR (CD₃OD): δ (ppm)2.90 (s, 3H), 3.02 (m, 4H), 3.38 (m, 4H), 4.61 (s, 2H).Methanesulfonylpiperazine was also prepared by reacting methanesulfonylchloride with excess piperazine (>2 equivalents) in water.

The racemic or single chiral isomer forms of 3-acetylaminopyrrolidinewere prepared by treating N¹-Boc-3-aminopyrrolidine (racemate, 3R, or3S) with acetyl chloride (iPr₂NEt, CH₂Cl₂, 0° C.), and deprotecting theN-Boc group (CF₃CO₂H, CH₂Cl₂). 3-(Acetamido)pyrrolidine: ¹H-NMR(DMSO-d₆; TFA salt): δ (ppm) 4.2 (quin, 1H), 3.3-3.1 (m, 3H), 2.9 (m,1H), 2.0 (m, 1H), 1.8 (br s, 4H).

3-((R)-2-Hydroxypropionamido)pyrrolidine was prepared after amidation ofN¹-Boc-3-aminopyrrolidine (L-lactic acid, PyBOP, DMF, RT), anddeprotection of N-Boc group (CF₃CO₂H, CH₂Cl₂). (m/z): [M+H]⁺ calcd forC₇H₁₄N₂O₂, 159.11; found, 159.0. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 4.4(quin, 1H), 4.1 (q, 1H), 3.5-3.4 (m, 2H), 3.3-3.2 (m, 2H), 2.3 (m, 1H),2.0 (m, 1H), 1.3 (d, 3H).

The N³-alkanesulfonyl derivatives of (3R)-aminopyrrolidine were obtainedby treating N¹-Boc-(3R)-aminopyrrolidine with propionylsulfonyl chlorideor cyclohexylmethylsulfonyl chloride (i-Pr₂NEt, CH₂Cl₂, 0° C.), anddeprotecting N-Boc group (CF₃CO₂H, CH₂Cl₂).

3-(N-Acetyl-N-methylamido)piperidine was prepared from N³-Cbz protected3-amino-piperidine-1-carboxylic acid t-butyl ester (De Costa, B., et al.J. Med. Chem. 1992, 35, 4334-43) after four synthetic steps: i) MeI,n-BuLi, THF, −78° C. to rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) AcCl,i-Pr₂NEt, CH₂Cl₂; iv) CF₃CO₂H, CH₂Cl₂. m/z: [M+H]⁺ calcd for C₈H₁₆N₂O:157.13; found, 157.2. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 4.6 (m, 1H), 3.3(m, 1H), 3.2 (m, 1H), 3.0 (m, 1H), 2.9 (s, 3H), 2.8 (m, 1H), 2.0 (s,3H), 1.9-1.7 (m, 4H).

3-(N-Acetyl-amido)piperidine was prepared from3-amino-piperidine-1-carboxylic acid tert-butyl ester afterN-acetylation and deprotection of the N-Boc group: i) AcCl, i-Pr₂NEt,CH₂Cl₂; ii) CF₃CO₂H, CH₂Cl₂. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 3.9 (m,1H), 3.3 (dd, 1H), 3.2 (m, 1H), 2.9 (dt, 1H), 2.75 (dt, 1H), 2.0-1.9 (m,2H), 1.9 (s, 3H), 1.8-1.4 (m, 2H).

The N³-alkanesulfonyl derivatives of 3-aminopiperidine were synthesizedby reacting the chiral or racemic forms of3-amino-piperidine-1-carboxylic acid tert-butyl ester with therespective alkanesulfonyl chloride (i-Pr₂NEt, CH₂Cl₂) and deprotectingthe N-Boc group (CF₃CO₂H, CH₂Cl₂).(3S)-3-(ethanesulfonylamido)piperidine: ¹H-NMR (CD₃OD): δ (ppm) 1.29 (t,3H, J₁=7.4 Hz), 1.50-1.80 (m, 2H), 1.90-2.10 (m, 2H), 2.89 (m, 2H), 3.05(q, 2H, J₁=7.4 Hz), 3.27 (m, 2H), 3.40 (d of d(br), 1H), 3.52 (m, 1H).3S-Methylsulfonylmethanesulfonylamido-piperidine: ¹H-NMR (CD₃OD): δ(ppm) 2.13-2.30 (m, 2H), 2.40-2.57 (m, 2H), 2.98 (m, 2H), 3.15 (s, 3H),3.21 (m, 2H), 3.30 (br d, 1H), 3.74 (m, 1H).

3-(Methylamino)-1-acetylpyrrolidine was prepared from3-(methylamino)-1-benzylpyrrolidine (TCI America) after four steps: i)(Boc)₂O, MeOH, rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) AcCl, i-Pr₂NEt,CH₂Cl₂; iv) CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd for C₇H₁₄N₂O: 143.12;found, 143.0.

3-(Methylamino)-1-(methanesulfonyl)pyrrolidine was prepared from3-(methylamino)-1-benzylpyrrolidine after four steps: i) (Boc)₂O, MeOH,rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) CH₃SO₂Cl, i-Pr₂NEt, CH₂Cl₂; iv)CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd for C₆H₁₄N₂O₂S: 179.08; found,179.2. 3R-Methylamino-1-(methanesulfonyl)pyrrolidine was prepared in asimilar manner from (3R)-(methylamino)-1-benzylpyrrolidine.

Derivatives of tetrahydro-3-thiophenamine-1,1-dioxide were preparedfollowing the protocol of Loev, B. J. Org. Chem. 1961, 26, 4394-9 byreacting 3-sulfolene with a requisite primary amine in methanol (cat.KOH, rt). N-Methyl-3-tetrahydrothiopheneamine-1,1-dioxide (TFA salt):¹H-NMR (DMSO-d₆): δ (ppm) 9.4 (br s, 2H), 4.0-3.8 (quin, 1H), 3.6-3.5(dd, 1H), 3.4-3.3 (m, 1H), 3.2-3.1 (m, 2H), 2.5 (s, 3H), 2.4 (m, 1H),2.1 (m, 1H).N-2-(1-hydroxy)ethyl-3-tetrahydrothiopheneamine-1,1-dioxide: (m/z):[M+H]⁺ calcd for C₆H₁₃NO₃S: 180.07; found, 180.2.

N-Methyl-tetrahydro-2H-thiopyran-4-amine-1,1-dioxide was prepared fromtetrahydro-4H-thiopyran-4-one: i) MeNH₂, NaBH₄; ii) (Boc)₂O, MeOH; iii)mCPBA, CH₂Cl₂, 0° C.; iv) CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd forC₆H₁₃NO₂S 164.07; found, 164.9. ¹H-NMR (CD₃OD; TFA salt): δ (ppm)3.4-3.1 (m, 5H), 2.7 (s, 3H), 2.4 (br d, 2H), 2.1 (br m, 2H).

1-Acetyl-3-(methylamino)piperidine was prepared from N³-Cbz protected3-methylamino-piperidine: i) AcCl, i-Pr₂NEt, CH₂Cl₂; ii) H₂ (1 atm), 10%Pd/C, EtOH. ¹H-NMR (CD₃OD): δ (ppm) 4.0 (m, 1H), 3.6 (m, 1H), 3.4-3.2(m, 2H), 3.0 (m, 1H), 2.6 (s, 3H), 2.1 (s, 3H), 1.8-1.6 (m, 4H).

1-(Methanesulfonyl)-3-(methylamino)piperidine was prepared from N³-Cbzprotected 3-methylamino-piperidine: i) CH₃SO₂Cl, i-Pr₂NEt, CH₂Cl₂; ii)H₂ (1 atm), 10% Pd/C, EtOH. (m/z): [M+H]⁺ calcd for C₇H₁₆N₂O₂S 193.10;found, 193.0. ¹H-NMR (DMSO-d₆; TFA salt): δ (ppm) 3.4 (dd, 1H), 3.2 (m,2H), 3.10 (s, 3H), 3.0-2.9 (m, 2H), 2.8 (s, 3H), 1.85-1.75 (m, 2H),1.6-1.4 (m, 2H).

Proline dimethylamide, and iminodiacetonitrile were purchased fromBachem, and Aldrich, respectively.

The N-derivatives of piperazine such as 1-(methoxycarbonyl)piperazine,1-(dimethylaminocarbonyl)piperazine, and1-(dimethylaminosulfonyl)piperazine were prepared by reacting piperazinewith methylchloroformate, dimethylaminochoroformate, ordimethylaminosulfamoyl chloride, respectively.

1-Methylamino-2-methylsulfonylethane was obtained by reactingmethylamine with methyl vinyl sulfone in methanol.N-[2-(2-methoxyethylamino)ethyl], N-methyl-methanesulfonamide wassynthesized starting from partially N-Boc protected ethanediamine afterfour steps of reactions in a sequence as follows: i) methylsulfonylchloride, triethylamine; ii) MeI, Cs₂CO₃; iii) NaH,1-bromo-2-methoxyethane; iv) CF₃CO₂H.

Isonipecotamide(piperidine-4-carboxamide), and proline amide werepurchased from Aldrich. 2-Hydroxymethylmorpholine was available fromTyger Scientific Product.

Methyl 4-piperidinylcarbamate was prepared from the reaction of N₁-Bocprotected 4-aminopiperidine with methylchloroformate followed by thedeprotection of the N-Boc group.

4-Piperidinol-dimethylcarbamate, and N-dimethyl-N′-(3-piperidinyl)ureawere prepared by reacting dimethylcarbamoyl chloride with N-Bocprotected 4-piperidinol or N₁-Boc-3-aminopiperidine, respectively.

3-(Methylamino)-1-(dimethylaminosulfonyl)pyrrolidine was obtained byreacting 3-(N-methyl-N-Boc-amino)pyrrolidine with dimethylsulfamoylchloride.

2-(3-Pyrrolidinyl)isothiazolidine-1,1-dioxide was synthesized bytreating N₁-Boc protected 3-aminopyrrolidine with 3-chloropropylsulfonylchloride in the presence of triethylamine, and followed by TFA treatmentfor the deprotection of the Boc group.

Example 1 Synthesis of1-Isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid(1S,3R,5R)-[8-(2-hydroxy-3-{[2-(2-hydroxyethoxy)ethyl]methanesulfonylamino}propyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

a. Preparation of 8-benzyl-8-azabicyclo[3.2.1]octan-3-one

Concentrated hydrochloric acid (30 mL) was added to a heterogeneoussolution of 2,5-dimethoxy tetrahydrofuran (82.2 g, 0.622 mol) in water(170 mL) while stirring. In a separate flask cooled to 0° C. (ice bath),concentrated hydrochloric acid (92 mL) was added slowly to a solution ofbenzyl amine (100 g, 0.933 mol) in water (350 mL). The2,5-dimethoxytetrahydrofuran solution was stirred for approximately 20min, diluted with water (250 mL), and then the benzyl amine solution wasadded, followed by the addition of a solution of 1,3-acetonedicarboxylicacid (100 g, 0.684 mol) in water (400 mL) and then the addition ofsodium hydrogen phosphate (44 g, 0.31 mol) in water (200 mL). The pH wasadjusted from pH 1 to pH ˜4.5 using 40% NaOH. The resulting cloudy andpale yellow solution was stirred overnight. The solution was thenacidified to pH 3 from pH 7.5 using 50% hydrochloric acid, heated to 85°C. and stirred for 2 hours. The solution was cooled to room temperature,basified to pH 12 using 40% NaOH, and extracted with DCM (3×500 mL). Thecombined organic layers were washed with brine, dried (MgSO₄), filteredand concentrated under reduced pressure to afford the crude titleintermediate as a viscous brown oil (52 g).

To a solution of the crude intermediate in methanol (1000 mL) was addeddi-tert-butyl dicarbonate (74.6 g, 0.342 mol) at 0° C. The solution wasallowed to warm to room temperature and stirred overnight. The methanolwas removed under reduced pressure and the resulting oil was dissolvedin dichloromethane (1000 mL). The intermediate was extracted into 1 MH₃PO₄ (1000 mL) and washed with dichloromethane (3×250 mL) The aqueouslayer was basified to pH 12 using aqueous NaOH, and extracted withdichloromethane (3×500 mL). The combined organic layers were dried(MgSO₄), filtered and concentrated under reduced pressure to produce thetitle intermediate as a viscous, light brown oil. ¹H-NMR (CDCl₃) δ (ppm)7.5-7.2 (m, 5H, C₆H₅), 3.7 (s, 2H, CH₂Ph), 3.45 (broad s, 2H, CH—NBn),2.7-2.6 (dd, 2H, CH₂CO), 2.2-2.1 (dd, 2H, CH₂CO), 2.1-2.0 (m, 2H,CH₂CH₂), 1.6 (m, 2H, CH₂CH₂). (m/z): [M+H]⁺ calcd for C₁₄H₁₇NO 216.14;found, 216.0.

b. Preparation of 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester

To a solution of 8-benzyl-8-azabicyclo[3.2.1]octan-3-one (75 g, 0.348mol) in EtOAc (300 mL) was added a solution of di-tert-butyl dicarbonate(83.6 g, 0.383 mol, 1.1 eq) in EtOAc (300 mL). The resulting solutionand rinse (100 mL EtOAc) was added to a 1 L Parr hydrogenation vesselcontaining 23 g of palladium hydroxide (20 wt. % Pd, dry basis, oncarbon, ˜50% wet with water; e.g. Pearlman's catalyst) under a stream ofnitrogen. The reaction vessel was degassed (alternating vacuum and N₂five times) and pressurized to 60 psi of H₂ gas. The reaction solutionwas agitated for two days and recharged with H₂ as needed to keep the H₂pressure at 60 psi until the reaction was complete as monitored bysilica thin layer chromatography. The solution was then filtered througha pad of Celite® and concentrated under reduced pressure to yield thetitle intermediate quantitatively as a viscous, yellow to orange oil. Itwas used in the next step without further treatment. ¹H NMR (CDCl₃)

(ppm) 4.5 (broad, 2H, CH—NBoc), 2.7 (broad, 2H, CH₂CO), 2.4-2.3 (dd, 2H,CH₂CH₂), 2.1 (broad m, 2H, CH₂CO), 1.7-1.6 (dd, 2H, CH₂CH₂), 1.5 (s, 9H,(CH₃)₃COCON)).

c. Preparation of(1S,3R,5R)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester

To a solution of the product of the previous step (75.4 g, 0.335 mol) inmethanol (1 L) was added ammonium formate (422.5 g, 6.7 mol), water (115mL) and 65 g of palladium on activated carbon (10% on dry basis, ˜50%wet with water; Degussa type E101NE/W) under a stream of N₂ whilestirring via mechanical stirrer. After 24 and 48 hours, additionalportions of ammonium formate (132 g, 2.1 mol) were added each time. Oncereaction progression ceased, as monitored by anal. HPLC, Celite® (>500g) was added and the resulting thick suspension was filtered and thenthe collected solid was rinsed with methanol (˜500 mL). The filtrateswere combined and concentrated under reduced pressure until all methanolhad been removed. The resulting cloudy, biphasic solution was thendiluted with 1M phosphoric acid to a final volume of ˜1.5 to 2.0 L at pH2 and washed with dichloromethane (3×700 mL). The aqueous layer wasbasified to pH 12 using 40% aq. NaOH, and extracted with dichloromethane(3×700 mL). The combined organic layers were dried over MgSO₄, filtered,and concentrated by rotary evaporation, then high-vacuum leaving 52 g(70%) of the title intermediate, commonly N-Boc-endo-3-aminotropane, asa white to pale yellow solid. The isomer ratio of endo to exo amine ofthe product was >99 based on ¹H-NMR analysis (>96% purity by analyticalHPLC). ¹H NMR (CDCl₃) δ (ppm) 4.2-4.0 (broad d, 2H, CHNBoc), 3.25 (t,1H, CHNH₂), 2.1-2.05 (m, 4H), 1.9 (m, 2H), 1.4 (s, 9H, (CH₃)₃OCON),1.2-1.1 (broad, 2H). (m/z): [M+H]⁺ calcd for C₁₂H₂₂N₂O₂) 227.18; found,227.2. Analytical HPLC (isocratic method; 2:98 (A:B) to 90:10 (A:B) over5 min): retention time=3.68 min.

d. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid

Acetone (228.2 mL, 3.11 mol) was added to a stirred suspension of2-aminophenylmethanol (255.2 g, 2.07 mol) and acetic acid (3.56 mL, 62mmol) in water (2 L) at room temperature. After 4 h, the suspension wascooled to 0° C. and stirred for an additional 2.5 h and then filtered.The solid was collected and washed with water and the wet solid cooledand dried by lyophilisation to yield2,2,-dimethyl-1,4-dihydro-2H-benzo[1,3]oxazine (332.2 g, 98%) as anoff-white solid. ¹H NMR (CDCl₃; 300 MHz): 1.48 (s, 6H, C(CH ₃)₂), 4.00(bs, 1H, NH), 4.86 (s, 2H, CH ₂), 6.66 (d, 1H, ArH), 6.81 (t, 1H, ArH),6.96 (d, 1H, ArH), 7.10 (t, 1H, ArH).

A solution of 2,2,-dimethyl-1,4-dihydro-2H-benzo[1,3]oxazine (125 g,0.77 mol) in THF (1 L) was filtered through a scintillation funnel andthen added dropwise via an addition funnel, over a period of 2.5 h, to astirred solution of 1.0 M LiAlH₄ in THF (800 mL) at 0° C. The reactionwas quenched by slow portionwise addition of Na₂SO₄ 10H₂O (110 g), overa period of 1.5 h, at 0° C. The reaction mixture was stirred overnight,filtered and the solid salts were washed thoroughly with THF. Thefiltrate was concentrated under reduced pressure to yield2-isopropylaminophenylmethanol (120 g, 95%) as a yellow oil. ¹H NMR(CDCl₃; 300 MHz): 1.24 (d, 6H, CH(CH ₃)₂), 3.15 (bs, 1H, OH), 3.61(sept, 1H, CH(CH₃)₂), 4.57 (s, 2H, CH ₂), 6.59 (t, 1H, ArH), 6.65 (d,1H, ArH), 6.99 (d, 1H, ArH), 7.15 (t, 1H, ArH).

Manganese dioxide (85% 182.6 g, 1.79 mol) was added to a stirredsolution of 2-isopropylaminophenylmethanol (118 g, 0.71 mol) in toluene(800 mL) and the reaction mixture was heated to 117° C. for 4 h. Thereaction mixture was allowed to cool to room temperature overnight andthen filtered through a pad of Celite which was eluted with toluene. Thefiltrate was concentrated under reduced pressure to yield2-isopropylaminobenzaldehyde (105 g, 90%) as an orange oil. ¹H NMR(CDCl₃; 300 MHz): 1.28 (d, 6H, CH(CH ₃)₂), 3.76 (sept, 1H, CH(CH₃)₂),6.65 (t, 1H, ArH), 6.69 (d, 1H, ArH), 7.37 (d, 1H, ArH), 7.44 (t, 1H,ArH), 9.79 (s, 1H, CHO).

2,2-Dimethyl-[1,3]dioxane-4,6-dione, commonly Meldrum's acid, (166.9 g,1.16 mol) was added to a stirred solution of2-isopropylaminobenzaldehyde (105 g, 0.64 mol), acetic acid (73.6 mL,1.29 mol) and ethylenediamine (43.0 mL, 0.64 mol) in methanol (1 L) at0° C. The reaction mixture was stirred for 1 h at 0° C. and then at roomtemperature overnight. The resulting suspension was filtered and thesolid washed with methanol and collected to yield the titleintermediate, 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid(146 g, 98%) as an off-white solid. ¹H NMR (CDCl₃; 300 MHz): 1.72 (d,6H, CH(CH ₃)₂), 5.50 (bs, 1H, CH(CH₃)₂), 7.44 (t, 1H, ArH), 7.75-7.77(m, 2H, ArH), 7.82 (d, 1H, ArH), 8.89 (s, 1H, CH).

e. Preparation of(1S,3R,5R)-3-[1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester

Thionyl chloride (36.6 mL, 0.52 mol) was added to a stirred suspensionof 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (80 g, 0.35mol) in toluene (600 mL) at 85° C. and the reaction mixture then heatedto 95° C. for 2 h. The reaction mixture was cooled to room temperatureand then added over 25 min to a vigorously stirred biphasic solution of(1S,3R,5R)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester (78.2 g, 0.35 mol) and sodium hydroxide (69.2 g, 1.73mol) in toluene/water (1:1) (1 L) at ° C. After 1 h, the layers wereallowed to separate and the organic phase concentrated under reducedpressure. The aqueous phase was washed with EtOAc (1 L) and then (500mL) and the combined organic extracts used to dissolve the concentratedorganic residue. This solution was washed with 1M H₃PO₄ (500 mL), sat.aq. NaHCO₃ (500 mL) and brine (500 mL), dried over MgSO₄, filtered andconcentrated under reduced pressure to yield the title intermediate(127.9 g, approx. 84%) as a yellow solid. ¹H NMR (CDCl₃): 1.47 (s, 9H),1.67 (d, 6H), 1.78-1.84 (m, 2H), 2.04-2.18 (m, 6H), 4.20-4.39 (m, 3H),5.65 (bs, 1H), 7.26 (dd. 1H), 7.63 (m, 2H), 7.75 (dd, 1H), 8.83 (s, 1H),10.63 (d, 1H).

f. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid {(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide

TFA (300 mL) was added to a stirred solution of the product of theprevious step (127.9 g) in CH₂Cl₂ (600 mL) at 0° C. The reaction mixturewas warmed to room temperature and stirred for 1 h and then concentratedunder reduced pressure. The oily brown residue was then poured into avigorously stirred solution of ether (3 L) and a solid precipitateformed immediately. The suspension was stirred overnight and then thesolid collected by filtration and washed with ether to yield the titleintermediate as its trifluoroacetic acid salt (131.7 g, 86% over twosteps) as a light yellow solid. ¹H NMR (CDCl₃): 1.68 (d, 6H), 2.10 (d,2H), 2.33-2.39 (m, 4H), 2.44-2.61 (m, 2H), 4.08 (bs, 2H), 4.41 (m, 1H),5.57 (bs, 1H), 7.31 (m. 1H), 7.66 (m, 2H), 7.77 (d, 1H), 8.83 (s, 1H),9.38 (bd, 2H), 10.78 (d, 1H).

g. Preparation of3-hydroxy-3′-{[1-isopropyl-2-oxo-1,2-dihydroquinolin-3-yl)carbonyl]amino}spiro[azetidine-1,8′-(1S,3R,5R)-8-azabicyclo[3.2.1]octane

(Intermediate (XV) with R¹=H, R²=isopropyl, R³=H)

2-Bromomethyloxirane (10.72 mL, 129.5 mmol) was added to a stirredsolution of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide trifluoroacetic acid salt(14.65 g, 43.2 mmol) in ethanol (150 mL) at room temperature. Thereaction mixture was stirred for 36 h, at which time a solid precipitateformed. The solid was collected by filtration and washed with ethanol(70 mL) to yield the title intermediate as the bromide salt (8.4 g).(m/z): [M+H]⁺ calcd for C₂₃H₃₀N₃O₃ 396.23; found, 396.5. Retention time(anal. HPLC: 2-50% MeCN/H₂O over 5 min)=4.13 min.

h. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid(1S,3R,5R)-8-{2-hydroxy-3-[2-(2-hydroxyethoxy)ethylamino]propyl}-8-azabicyclo[3.2.1]oct-3-yl)-amide

(Intermediate (III) with R¹=H, R²=isopropyl, R³=H, n=1,Y=2-hydroxyethoxy)

3-Hydroxy-3′-{[1-isopropyl-2-oxo-1,2-dihydroquinolin-3-yl)carbonyl]amino}-spiro[azetidine-1,8′-(1S,3R,5R)-8-azabicyclo[3.2.1]octane,the product of step (g), (115 mg, 0.24 mmol), was added to 1.0 mLethanol, 2-(2-aminoethoxy)ethanol (50.46 mg, 0.48 mmol) andN,N-diisopropylethylamine (93.06 mg, 0.72 mmol). The resulting mixturewas heated to 80° C. overnight, then concentrated under reduced pressureto provide the title compound as an intermediate of formula (III) whichwas used without further isolation or purification.

i. Synthesis of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(2-hydroxy-3-{[2-(2-hydroxyethoxy)ethyl]methanesulfonylamino}propyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

The product of step (h) above was dissolved in dichloromethane (1.0 mL).N,N-diisopropylethylamine (62.04 mg, 0.48 mmol) was added to themixture, and the mixture was then cooled to 0° C. To this cold mixturewas slowly added methanesulfonylchloride (93.06 mg, 0.528 mmol). Afterstirring at 0° C. for one hour, the reaction was quenched with 0.5 mL50% CH₃CO₂H in H₂O, concentrated, redissolved in 1.5 mL 50% CH₃CO₂H inH₂O, and purified by reversed phase preparative HPLC to yield the titlecompound as trifluoroacetic acid salt. (m/z): [M+H]⁺ calcd forC₂₈H₄₂N₄O₇S 579.29; found 579.2.

Example 2 Synthesis of1-Isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{(1S,3R,5R)-8-[3-(2-ethylsulfanylethylamino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}-amide(Intermediate (III) with R¹=H, R²=isopropyl, R³=H, n=1, Y=—SCH₂CH₃)

Using the process and reagents described in Example 1, step (h) above,except substituting (ethylthio)ethylamine for 2-(2-aminoethoxy)ethanol,the title intermediate of formula (III) was prepared. After the reactionmixture was concentrated, the residue was diluted with 1.5 mL 50%CH₃CO₂H in H₂O and purified by reversed phase preparative HPLC.

b. Synthesis of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-azabicyclo-[3.2.1]oct-3-yl)amide

A dichloromethane solution of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU)(0.668 M, 67 μL was added to the product of the previous step (11.1 mg,0.015 mmol). The mixture was cooled to 0° C. before a solution ofmethanesulfonylchloride in dichloromethane (1.268 M, 11.8 μL) was added.The reaction progress was monitored by mass spectrum for approximatelyone hour until the desired product was observed together with startingmaterial. Another 11.8 μL (1.268M) dichloromethane solution ofmethanesulfonylchloride was added. The mixture was stirred at 0° C. foranother hour. Then the reaction was quenched with a mixture of 50%CH₃COOH in water, concentrated, redissolved in 50% CH₃COOH in water, andpurified by reversed phase preparative HPLC to yield the title compoundas a trifluoroacetic acid salt. (m/z): [M+H]⁺ calcd for C₂₈H₄₂N₄O₅S₂579.27; found 579.2.

Example 3 Synthesis of(2-Hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(4-methanesulfonylpiperazin-1-yl)ethyl]-carbamicacid methyl ester

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{8-[3-(2,2-dimethoxyethylamino)-2-hydroxy-propyl]-8-(1S,3R,5R)-azabicyclo[3.2.1]oct-3-yl}amide

To a stirred solution of3-hydroxy-3′-{[1-isopropyl-2-oxo-1,2-dihydroquinolin-3-yl)carbonyl]amino}spiro[azetidine-1,8′-(1S,3R,5R)-8-azabicyclo[3.2.1]octane,the product of step (g) of Example 1, (6.8 g, 14.27 mmol) in ethanol(150 mL) was added DIPEA (3.69 g, 28.55 mmol) and2,2-dimethoxyethylamine (4.5 g, 42.82 mmol). The reaction was heated toreflux and stirred overnight. The reaction solution was allowed to coolto ambient temperature and was then concentrated under reduced pressure.The concentrate was diluted with DCM (500 mL) and then extracted with 1MH₃PO₄ (500 mL). The aqueous phase was washed with DCM (2×200 mL),basified to pH=12 (40% NaOH) and extracted with DCM (3×300 mL). Theorganic phase was dried (MgSO₄), filtered and concentrated under reducedpressure to yield the title intermediate (6.5 g) as a light brown,viscous oil. (m/z): [M+H]⁺ calcd for C₂₇H₄₀N₄O₅ 501.30; found 501.6.

b. Preparation of(2,2-Dimethoxyethyl)-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamicacid methyl ester

To a stirred solution of the product of the previous step (2.16 g, 4.315mmol) in DCM (50 mL) was added DIPEA (0.585 g, 4.531 mmol). The reactionwas cooled to 0° C. and methylchloroformate (0.428 g, 4.531 mmol) wasadded dropwise. The stirred reaction was allowed to warm to roomtemperature overnight. The reaction solution was concentrated underreduced pressure, dissolved in 50% aqueous acetonitrile (10 mL), andpurified by preparative HPLC. The clean fractions were combined andlyophilized to yield the title intermediate (1.2 g) as a white solid.(m/z): [M+H]⁺ calcd for C₂₉H₄₂N₄O₇ 559.31; found 559.6.

c. Preparation of(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-(2-oxoethyl)-carbamicacid methyl ester (Intermediate (V) with R¹=H, R²=isopropyl, R³=H, andR⁴ is —C(O)OCH₃)

To the product of the previous step (1.1 g, 1.635 mmol) was added 50%HCl (10 mL). The reaction solution was stirred for 30 minutes, and thenexcess HCl was evaporated under reduced pressure. The resulting solutionwas diluted with 50% aqueous acetonitrile and lyophilized, to yield thetitle intermediate (0.875 g) as a pale yellow solid. (m/z): [M+H]⁺ calcdfor C₂₇H₃₆N₄O₆ 513.26; found 513.4.

d. Synthesis of(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(4-methanesulfonyl-piperazin-1-yl)ethyl]-carbamicacid methyl ester

To a solution of piperazinesulfonamide (25.5 mg, 0.0918 mmol) and DIPEA(71 mg, 0.55 mmol) in DCM (0.5 mL) was added a solution of the productof the previous step (0.017 g, 0.031 mmol) in DCM (0.5 mL), and thesolution was agitated for 5 minutes. Sodium triacetoxyborohydride(0.0091 g, 0.043 mmol) was added and the resulting suspension wasagitated for 1 hour. The solution was concentrated under reducedpressure to dryness, then dissolved in 50% aqueous acetonitrile andpurified by HPLC. The purified fractions were combined and lyophilizedto yield the title compound (0.03 g) as a trifluoroacetate salt. (m/z):[M+H]⁺ calcd for C₃₂H₄₈N₆O₇S 661.34; found, 661.2. Retention time (anal.HPLC: 5-60% MeCN/H₂O over 5 min)=2.11 min.

Example 4 Synthesis of1-Isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(acetyl-{2-[(2-methanesulfonylethyl)-methylamino]-ethyl}amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid((1S,3R,5R)-8-{3-[acetyl-(2,2-dimethoxyethyl)amino]-2-hydroxypropyl}-8-azabicyclo-[3.2.1]oct-3-yl)-amide

Using the synthetic process and reagents described in Example 3, Step b,except substituting acetyl chloride (0.355 g, 4.531 mmol) formethylchloroformate, the title intermediate (1.2 g) was prepared as awhite solid. (m/z): [M+H]⁺ calcd for C₂₉H₄₂N₄O₆ 543.31; found 543.8.

b. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid((1S,3R,5R)-8-{3-[acetyl-(2-oxo-ethyl)-amino]-2-hydroxy-propyl}-8-aza-bicyclo[3.2.1]oct-3-yl)-amide(Intermediate (V) with R¹=H, R²=isopropyl, R³=H, and R⁴ is —C(O)CH₃)

To the product of the previous step (1.0 g, 1.523 mmol) was added 50%HCl (10 mL). The reaction solution was stirred for 30 minutes, and thenexcess HCl was evaporated at reduced pressure. The resulting solutionwas diluted with 50% aqueous acetonitrile and lyophilized, to yield thetitle intermediate (0.991 g) as a pale yellow solid. (m/z): [M+H]⁺ calcdfor C₂₇H₃₆N₄O₅ 497.27; found 497.6.

c. Synthesis of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(acetyl-{2-[(2-methanesulfonylethyl)-methylamino]-ethyl}amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

To a solution of (2-methanesulfonylethyl)methylamine (13.7 mg, 0.103mmol) and DIPEA (27.1 mg, 0.21 mmol) in DCM (0.5 mL) was added asolution of the product of the previous step (0.019 g, 0.035 mmol) inDCM (0.5 mL). The solution was agitated for 5 minutes. Sodiumtriacetoxyborohydride (0.010 g, 0.048 mmol) was added and the resultingsuspension was agitated for 1 hour. The solution was concentrated underreduced pressure to dryness, then dissolved in 50% aqueous acetonitrileand purified by HPLC. The purified fractions were combined andlyophilized to yield the title compound (0.005 g) as a trifluoroacetatesalt. (m/z): [M+H]⁺ calcd for C₃₁H₄₇N₅O₆S 618.33; found, 618.2.Retention time (anal. HPLC: 5-60% MeCN/H₂O over 5 min)=2.01 min.

Using the methods described in Examples 1-4, and substituting theappropriate reagents, the following compounds listed in Tables 1-6 wereprepared. In all of the compounds of the invention depicted in Tables1-6, the quinolinone-carboxamide is endo to the azabicyclooctane group.

TABLE 1 (II-a)

Molecular Calc'd Obsd # n R⁴ Y Formula [M + H] [M + H] 1 1 —SO₂CH₃—OCH₂CH₂OH C₂₈H₄₂N₄O₇S 575.29 575.2 2 1 —SO₂CH₃ —SCH₂CH₃ C₂₈H₄₂N₄O₅S₂579.27 579.2 3 0 —SO₂CH₃

C₂₉H₄₂N₄O₆S 575.29 575.2 4 2 —SO₂CH₃

C₃₀H₄₂N₆O₅S 599.30 599.2 5 1 —SO₂CH₃

C₃₄H₄₃N₅O₅S 634.31 634.2 6 1 —SO₂CH₃ —NHSO₂CH₃ C₂₇H₄₁N₅O₇S₂ 612.25 612.27 2 —SO₂CH₃

C₃₁H₄₅N₅O₆S 616.32 616.2 8 0 —SO₂CH₃

C₂₉H₄₂N₄O₆S 575.29 575.2 9 1 —SO₂CH₃

C₃₁N₄₁N₅O₅S 596.29 596.2 10 1 —SO₂CH₃ —OCH3 C₂₇H₄₀N₄O₆S 549.71 549.2 111 —C(O)OCH₃

C₃₃H₅₀N₆O₇S 675.36 675.2 12 1 —C(O)OCH₃

C₃₂H₄₆N₆O₆ 611.36 611.2 13 1 —C(O)OCH₃ —N(CH₃)SO₂N(CH₃)₂ C₃₃H₅₃N₇O₇S692.38 692.2 14 1 —C(O)OCH₃ —N(CH₂CH₂OCH₃)—(CH₂)₂N(CH₃)SO₂CH₃C₃₄H₅₄N₆O₈S 707.38 707.2 15 1 —C(O)OCH₃ —N(CH₃)CH₂—C(O)N(CH₃)₂C₃₂H₄₈N₆O6 613.37 613.2 16 1 —C(O)OCH₃ —N(CH₃)CH₂C(O)NH₂ C₃₀H₄₄N₆O₆585.34 585.2 17 1 —C(O)CH₃

C₃₄H₅₂N₆O₆S 673.38 673.2 18 1 —C(O)CH₃ —N(CH₃)CH₂CH₂N(CH₃)SO₂CH₃C₃₂H₅₀N₆O₆S 647.36 647.2 19 1 —C(O)CH₃ —N(CH₃)CH₂CH₂NHSO₂CH₃ C₃₁H₄₈N₆O₆S633.35 633.2 20 1 —C(O)CH₃ —N(CH₃)CH₂CH₂N(CH₃)C(O)OCH₃ C₃₃H₅₀N₆O₆ 627.39627.2 21 1 —C(O)CH₃ —N(CH₃)CH₂CH₂N(CH₃)SO₂N(CH₃)₂ C₃₃H₅₃N₇O₆S 676.39676.2 22 1 —C(O)CH₃ —N(CH₃)CH₂CH₂SO₂CH₃ C₃₁H₄₇N₅O₆S 618.33 618.2 23 1—C(O)CH₃ —N(CH₂CH₂OCH₃)—(CH₂)₂SO₂CH₃ C₃₃H₅₁N₅O₇S 662.36 662.2 24 1—C(O)CH₃ —N(CH₂CH₂OCH₃)—(CH₂)₂N(CH₃)SO₂CH₃ C₃₄H₅₄N₆O₇S 691.39 691.3 25 1—C(O)CH₃ —N(CH₃)CH₂C(O)N(CH₃)₂ C₃₂H₄₈N₆O₅ 597.38 597.3 26 1 —C(O)CH₃—N(CH₃)CH₂C(O)NH₂ C₃₀H₄₄N₆O₅ 569.35 569.2 27 1 —C(O)CH₃

C₃₃H₅₀N₆O₆S 659.36 659.2

TABLE 2 (II-b)

Molecular Calc'd Obsd # R⁴ Z Formula [M + H] [M + H] 1 —C(O)OCH₃3-N(CH₃)SO₂CH₃ C₃₄H₅₂N₆O₇S 689.37 689.2 2 —C(O)OCH₃ 3-N(CH₃)C(O)OCH₃C₃₅H₅₂N₆O₇ 669.4 669.3 3 —C(O)OCH₃ 3-N(CH₃)C(O)CH₃ C₃₅H₅₂N₆O₆ 653.4653.3 4 —C(O)OCH₃ 3-NHSO₂N(CH₃)₂ C₃₄H₅₃N₇O₇S 704.38 704.2 5 —C(O)OCH₃3-NHC(O)OCH₃ C₃₄H₅₀N₆O₇ 655.38 655.2 6 —C(O)OCH₃ 3-NHC(O)CH₃ C₃₄H₅₀N₆O₆639.39 639.3 7 —C(O)OCH₃ 3-NHC(O)N(CH₃)₂ C₃₅H₅₃N₇O₆ 668.42 668.3 8—C(O)OCH₃ 3-C(O)NH₂ C₃₃H₄₈N₆O₆ 625.37 625.2 9 —C(O)OCH₃ 4-OC(O)N(CH₃)₂C₃₅H₅₂N₆O₇ 669.4 669.2 10 —C(O)CH₃ 3-N(CH₃)SO₂CH₃ C₃₄H₅₂N₆O₆S 673.38673.2 11 —C(O)CH₃ 3-N(CH₃)C(O)OCH₃ C₃₅H₅₂N₆O₆ 653.4 653.3 12 —C(O)CH₃3-N(CH₃)C(O)CH₃ C₃₅H₅₂N₆O₅ 637.41 637.3 13 —C(O)CH₃ 3-NHSO₂N(CH₃)₂C₃₄H₅₃N₇O₆S 688.39 688.2 14 —C(O)CH₃ 3-NHC(O)CH₃ C₃₄H₅₀N₆O₅ 623.39 623.315 —C(O)CH₃ 3-C(O)NH₂ C₃₃H₄₈N₆O₅ 609.38 609.2 16 —C(O)CH₃ 4-OC(O)N(CH₃)₂C₃₅H₅₂N₆O₆ 653.4 653.3

TABLE 3 (II-c)

Molecular Calc'd Obsd # R⁴ (CR⁶R⁷)m Z′ Formula [M + H] [M + H] 1—C(O)OCH₃ —CH(CH₂OH)CH₂— O C₃₂H₄₇N₅O₇ 614.36 614.3 2 —C(O)OCH₃ —(CH₂)₂—NSO₂CH₃ C₃₂H₄₈N₆O₇S 661.34 661.2 3 —C(O)OCH₃ —(CH₂)₂— NC(O)N(CH₃)₂C₃₄H₅₁N₇O₆ 654.4 654.3 4 —C(O)OCH₃ —(CH₂)₃— NC(O)CH₃ C₃₄H₅₀N₆O₆ 639.39639.3 5 —C(O)OCH₃ —(CH₂)₂— NC(O)NHCH₃ C₃₃H₄₉N₇O₆ 640.38 640.2 6—C(O)OCH₃ —(CH₂)₂— SO₂ C₃₁H₄₅N₅O₇S 632.31 632.3 7 —C(O)OCH₃ —(CH₂)₂—NC(O)NH₂ C₃₂H₄₇N₇O₆ 626.37 626.2 8 —C(O)CH₃ —CH(CH₂OH)CH₂— O C₃₂H₄₇N₅O₆598.36 598.2 9 —C(O)CH₃ —(CH₂)₂— NC(O)N(CH₃)₂ C₃₄H₅₁N₇O₅ 638.41 638.2 10—C(O)CH₃ —(CH₂)₃— NSO₂CH₃ C₃₃H₅₀N₆O₆S 659.36 659.2 11 —C(O)CH₃ —(CH₂)₃—NC(O)CH₃ C₃₄H₅₀N₆O₅ 623.39 623.3 12 —C(O)CH₃ —(CH₂)₂— NC(O)NH₂C₃₂H₄₇N₇O₅ 610.37 610.2 13 —C(O)CH₃ —(CH₂)₂— SO₂ C₃₁H₄₅N₅O₆S 616.32616.3

TABLE 4 (II-d)

Molecular Calc'd Obsd # R⁴ R⁵ Z′ Formula [M + H] [M + H] 1 —C(O)OCH₃—CH₃ SO₂ C₃₃H₄₉N₅O₇S 660.35 660.2 2 —C(O)CH₃ —CH₃ SO₂ C₃₃H₄₉N₅O₆S 644.35644.2

TABLE 5 (II-e)

Molecular Calc'd Obsd # R⁴ Z Formula [M + H] [M + H] 1 —C(O)OCH₃—NHC(O)CH₃ C₃₃H₄₈N₆O₆ 625.37 625.2 2 —C(O)OCH₃ —NHC(O)OCH₃ C₃₃H₄₈N₆O₇641 37 641.2 3 —C(O)OCH₃ —NHSO₂N(CH₃)₂ C₃₃H₅₁N₇O₇S 690.37 690.2 4—C(O)OCH₃ —OC(O)N(CH₃)₂ C₃₄H₅₀N₆O₇ 655.38 655.2 5 —C(O)OCH₃—N(CH₃)C(O)CH₃ C₃₄H₅₀N₆O₆ 639.39 639.2 6 —C(O)OCH₃ —N(CH₃)C(O)OCH₃C₃₄H₅₀N₆O₇ 655.38 655.3 7 —C(O)OCH₃ —N(CH₃)SO₂N(CH₃)2 C₃₄H₅₃N₇O₇S 704.38704.2 8 —C(O)OCH₃

C₃₄H₅₀N₆O₇S 687.36 687.3 9 —C(O)CH₃ —NHC(O)CH₃ C₃₃H₄₈N₆O₅ 609.38 609.210 —C(O)CH₃ —NHC(O)OCH₃ C₃₃H₄₈N₆O₆ 625.37 625.2 11 —C(O)CH₃—NHSO₂N(CH₃)₂ C₃₃H₅₁N₇O₆S 674.37 674.2 12 —C(O)CH₃ —OC(O)N(CH₃)₂C₃₄H₅₀N₆O₆ 639.39 639.3 13 —C(O)CH₃ —N(CH₃)C(O)CH₃ C₃₄H₅₀N₆O₅ 623.39623.3 14 —C(O)CH₃ —N(CH₃)C(O)OCH₃ C₃₄H₅₀N₆O₆ 639.39 639.2 15 —C(O)CH₃—N(CH₃)SO₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.39 688.2 16 —C(O)CH₃—N(CH₃)C(O)N(CH₃)₂ C₃₅H₅₃N₇O₅ 652.42 652.3

TABLE 6 (II-f)

Molecular Calc'd Obsd # R⁴ R⁵ Z′ Formula [M + H] [M + H] 1 —C(O)OCH₃—CH₃ NSO₂CH₃ C₃₃H₅₀N₆O₇S 675.36 675.2 2 —C(O)OCH₃ —CH₃ NSO₂N(CH₃)₂C₃₄H₅₃N₇O₇S 704.38 704.2 3 —C(O)OCH₃ —CH₃ NC(O)OCH₃ C₃₄H₅₀N₆O₇ 655.38655.3 4 —C(O)CH₃ —CH₃ NSO₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.39 688.3 5 —C(O)CH₃—CH₃ NC(O)OCH₃ C₃₄H₅₀N₆O₆ 639.39 639.3

Example 5 Radioligand Binding Assay on 5-HT_(4(c)) Human Receptors

a. Membrane Preparation 5-HT_(4(c))

HEK-293 (human embryonic kidney) cells stably-transfected with human5-HT_(4(c)) receptor cDNA (Bmax=˜6.0 μmol/mg protein, as determinedusing [³H]-GR113808 membrane radioligand binding assay) were grown inT-225 flasks in Dulbecco's Modified Eagles Medium (DMEM) containing4,500 mg/L D-glucose and pyridoxine hydrochloride (GIBCO-InvitrogenCorp., Carlsbad Calif.: Cat #11965) supplemented with 10% fetal bovineserum (FBS) (GIBCO-Invitrogen Corp.: Cat #10437), 2 mM L-glutamine and(100 units) penicillin-(100 μg) streptomycin/ml (GIBCO-Invitrogen Corp.:Cat #15140) in a 5% CO₂, humidified incubator at 37° C. Cells were grownunder continuous selection pressure by the addition of 800 μg/mLgeneticin (GIBCO-Invitrogen Corp.: Cat #10131) to the medium.

Cells were grown to roughly 60-80% confluency (<35 subculture passages).At 20-22 hours prior to harvesting, cells were washed twice and fed withserum-free DMEM. All steps of the membrane preparation were performed onice. The cell monolayer was lifted by gentle mechanical agitation andtrituration with a 25 mL pipette. Cells were collected by centrifugationat 1000 rpm (5 min).

For the membrane preparation, cell pellets were resuspended in ice-cold50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES), pH7.4 (membrane preparation buffer) (40 mL/total cell yield from 30-40T225 flasks) and homogenized using a polytron disrupter (setting 19,2×10 s) on ice. The resultant homogenates were centrifuged at 1200 g for5 min at 4° C. The pellet was discarded and the supernatant centrifugedat 40,000 g (20 min). The pellet was washed once by resuspension withmembrane preparation buffer and centrifugation at 40,000 g (20 min). Thefinal pellet was resuspended in 50 mM HEPES, pH 7.4 (assay buffer)(equivalent 1 T225 flask/1 mL). Protein concentration of the membranesuspension was determined by the method of Bradford (Bradford, 1976).Membranes were stored frozen in aliquots at −80° C.

b. Radioligand Binding Assays

Radioligand binding assays were performed in 1.1 mL 96-deep wellpolypropylene assay plates (Axygen) in a total assay volume of 400 μLcontaining 2 μg membrane protein in 50 mM HEPES pH 7.4, containing0.025% bovine serum albumin (BSA). Saturation binding studies fordetermination of K_(d) values of the radioligand were performed using[³H]-GR113808 (Amersham Inc., Bucks, UK: Cat #TRK944; specific activity˜82 Ci/mmol) at 8-12 different concentrations ranging from 0.001 mM-5.0nM. Displacement assays for determination of pK_(i) values of compoundswere performed with [³H]-GR113808 at 0.15 nM and eleven differentconcentrations of compound ranging from 10 μM-100 μM.

Test compounds were received as 10 mM stock solutions in DMSO anddiluted to 400 μM into 50 mM HEPES pH 7.4 at 25° C., containing 0.1%BSA, and serial dilutions (1:5) then made in the same buffer.Non-specific binding was determined in the presence of 1 μM unlabeledGR113808. Assays were incubated for 60 min at room temperature, and thenthe binding reactions were terminated by rapid filtration over 96-wellGF/B glass fiber filter plates (Packard BioScience Co., Meriden, Conn.)presoaked in 0.3% polyethyleneimine. Filter plates were washed threetimes with filtration buffer (ice-cold 50 mM HEPES, pH7.4) to removeunbound radioactivity. Plates were dried, 35 μL Microscint-20 liquidscintillation fluid (Packard BioScience Co., Meriden, Conn.) was addedto each well and plates were counted in a Packard Topcount liquidscintillation counter (Packard BioScience Co., Meriden, Conn.).

Binding data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 3-parameter model for one-site competition. The BOTTOM(curve minimum) was fixed to the value for nonspecific binding, asdetermined in the presence of 1 μM GR113808. K_(i) values for testcompounds were calculated, in Prism, from the best-fit IC₅₀ values, andthe K_(d) value of the radioligand, using the Cheng-Prusoff equation(Cheng and Prusoff, Biochemical Pharmacology, 1973, 22, 3099-108):K_(i)=IC₅₀/(1+[L]/K_(d)) where [L]=concentration [³H]-GR113808. Resultsare expressed as the negative decadic logarithm of the K_(i) values,pK_(i).

Test compounds having a higher pK_(i) value in this assay have a higherbinding affinity for the 5-HT₄ receptor. The compounds of the inventionwhich were tested in this assay had a pK_(i) value ranging from about6.3 to about 8.5, typically ranging from about 7.0 to about 8.0.

Example 6 Radioligand Binding Assay on 5-HT_(3A) Human Receptors:Determination of Receptor Subtype Selectivity

a. Membrane Preparation 5-HT_(3A)

HEK-293 (human embryonic kidney) cells stably-transfected with human5-HT_(3A) receptor cDNA were obtained from Dr. Michael Bruess(University of Bonn, GDR) (Bmax=˜9.0 pmol/mg protein, as determinedusing [³H]-GR65630 membrane radioligand binding assay). Cells were grownin T-225 flasks or cell factories in 50% Dulbecco's Modified EaglesMedium (DMEM) (GIBCO-Invitrogen Corp., Carlsbad, Calif.: Cat #11965) and50% Ham's F12 (GIBCO-Invitrogen Corp.: Cat #11765) supplemented with 10%heat inactivated fetal bovine serum (FBS) (Hyclone, Logan, Utah: Cat#SH30070.03) and (50 units) penicillin-(50 μg) streptomycin/ml(GIBCO-Invitrogen Corp.: Cat #15140) in a 5% CO₂, humidified incubatorat 37° C.

Cells were grown to roughly 70-80% confluency (<35 subculture passages).All steps of the membrane preparation were performed on ice. To harvestthe cells, the media was aspirated and cells were rinsed with Ca²⁺,Mg²⁺-free Dulbecco's phosphate buffered saline (dPBS). The cellmonolayer was lifted by gentle mechanical agitation. Cells werecollected by centrifugation at 1000 rpm (5 min). Subsequent steps of themembrane preparation followed the protocol described above for themembranes expressing 5-HT_(4(c)) receptors.

b. Radioligand Binding Assays

Radioligand binding assays were performed in 96-well polypropylene assayplates in a total assay volume of 200 μL containing 1.5-2 μg membraneprotein in 50 mM HEPES pH 7.4, containing 0.025% BSA assay buffer.Saturation binding studies for determination of K_(d) values of theradioligand were performed using [³H]-GR65630 (PerkinElmer Life SciencesInc., Boston, Mass.: Cat #NET1101, specific activity ˜85 Ci/mmol) attwelve different concentrations ranging from 0.005 nM to 20 nM.Displacement assays for determination of pK_(i) values of compounds wereperformed with [³H]-GR65630 at 0.50 nM and eleven differentconcentrations of compound ranging from 10 pM to 100 μM. Compounds werereceived as 10 mM stock solutions in DMSO (see section 3.1), diluted to400 μM into 50 mM HEPES pH 7.4 at 25° C., containing 0.1% BSA, andserial (1:5) dilutions then made in the same buffer. Non-specificbinding was determined in the presence of 10 μM unlabeled MDL72222.Assays were incubated for 60 min at room temperature, then the bindingreactions were terminated by rapid filtration over 96-well GF/B glassfiber filter plates (Packard BioScience Co., Meriden, Conn.) presoakedin 0.3% polyethyleneimine. Filter plates were washed three times withfiltration buffer (ice-cold 50 mM HEPES, pH7.4) to remove unboundradioactivity. Plates were dried, 35 μL Microscint-20 liquidscintillation fluid (Packard BioScience Co., Meriden, Conn.) was addedto each well and plates were counted in a Packard Topcount liquidscintillation counter (Packard BioScience Co., Meriden, Conn.).

Binding data were analyzed using the non-linear regression proceduredescribed above to determine K_(i) values. The BOTTOM (curve minimum)was fixed to the value for nonspecific binding, as determined in thepresence of 10 μM MDL72222. The quantity [L] in the Cheng-Prusoffequation was defined as the concentration [³H]-GR65630.

Selectivity for the 5-HT₄ receptor subtype with respect to the 5-HT₃receptor subtype was calculated as the ratioK_(i)(5-HT_(3A))/K_(i)(5-HT_(4(c))). The compounds of the inventionwhich were tested in this assay had a 5-HT₄/5-HT₃ receptor subtypeselectivity ranging from about 50 to about 8000, typically ranging fromabout 100 to about 4000.

Example 7 Whole-Cell cAMP Accumulation Flashplate Assay with HEK-293Cells Expressing Human 5-HT_(4(c)) Receptors

In this assay, the functional potency of a test compound was determinedby measuring the amount of cyclic AMP produced when HEK-293 cellsexpressing 5-HT₄ receptors were contacted with different concentrationsof test compound.

a. Cell Culture.

HEK-293 (human embryonic kidney) cells stably-transfected with clonedhuman 5-HT_(4(c)) receptor cDNA were prepared expressing the receptor attwo different densities: (1) at a density of about 0.5-0.6 pmol/mgprotein, as determined using a [³H]-GR113808 membrane radioligandbinding assay, and (2) at a density of about 6.0 μmol/mg protein. Thecells were grown in T-225 flasks in Dulbecco's Modified Eagles Medium(DMEM) containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat#11965) supplemented with 10% fetal bovine serum (FBS) (GIBCO-InvitrogenCorp.: Cat #10437) and (100 units) penicillin-(100 μg) streptomycin/ml(GIBCO-Invitrogen Corp.: Cat #15140) in a 5% CO₂, humidified incubatorat 37° C. Cells were grown under continuous selection pressure by theaddition of geneticin (800 μg/mL: GIBCO-Invitrogen Corp.: Cat #10131) tothe medium.

b. Cell Preparation

Cells were grown to roughly 60-80% confluency. Twenty to twenty-twohours prior to assay, cells were washed twice, and fed, with serum-freeDMEM containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat#11965). To harvest the cells, the media was aspirated and 10 mL Versene(GIBCO-Invitrogen Corp.: Cat #15040) was added to each T-225 flask.Cells were incubated for 5 min at RT and then dislodged from the flaskby mechanical agitation. The cell suspension was transferred to acentrifuge tube containing an equal volume of pre-warmed (37° C.) dPBSand centrifuged for 5 min at 1000 rpm. The supernatant was discarded andthe pellet was re-suspended in pre-warmed (37° C.) stimulation buffer(10 mL equivalent per 2-3 T-225 flasks). This time was noted and markedas time zero. The cells were counted with a Coulter counter (count above8 μm, flask yield was 1-2×10⁷ cells/flask). Cells were resuspended at aconcentration of 5×10⁵ cells/ml in pre-warmed (37° C.) stimulationbuffer (as provided in the flashplate kit) and preincubated at 37° C.for 10 min.

cAMP assays were performed in a radioimmunoassay format using theFlashplate Adenylyl Cyclase Activation Assay System with ¹²⁵I-cAMP(SMP004B, PerkinElmer Life Sciences Inc., Boston, Mass.), according tothe manufacturer's instructions.

Cells were grown and prepared as described above. Final cellconcentrations in the assay were 25×10³ cells/well and the final assayvolume was 100 μL. Test compounds were received as 10 mM stock solutionsin DMSO, diluted to 400 μM into 50 mM HEPES pH 7.4 at 25° C., containing0.1% BSA, and serial (1:5) dilutions then made in the same buffer.Cyclic AMP accumulation assays were performed with 11 differentconcentrations of compound ranging from 10 μM to 100 μM (final assayconcentrations). A 5-HT concentration-response curve (10 μM to 100 μM)was included on every plate. The cells were incubated, with shaking, at37° C. for 15 min and the reaction terminated by addition of 100 μl ofice-cold detection buffer (as provided in the flashplate kit) to eachwell. The plates were sealed and incubated at 4° C. overnight. Boundradioactivity was quantified by scintillation proximity spectroscopyusing the Topcount (Packard BioScience Co., Meriden, Conn.).

The amount of cAMP produced per mL of reaction was extrapolated from thecAMP standard curve, according to the instructions provided in themanufacturer's user manual. Data were analyzed by nonlinear regressionanalysis with the GraphPad Prism Software package using the 3-parametersigmoidal dose-response model (slope constrained to unity). Potency dataare reported as pEC₅₀ values, the negative decadic logarithm of the EC₅₀value, where EC₅₀ is the effective concentration for a 50% maximalresponse.

Test compounds exhibiting a higher pEC₅₀ value in this assay have ahigher potency for agonizing the 5-HT₄ receptor. The compounds of theinvention which were tested in this assay, for example, in the cell line(1) having a density of about 0.5-0.6 pmol/mg protein, had a pEC₅₀ valueranging from about 7.0 to about 9.0, typically ranging from about 7.5 toabout 8.5.

Example 8 In Vitro Voltage Clamp Assay of Inhibition of Potassium IonCurrent in Whole Cells Expressing the hERG Cardiac Potassium Channel

CHO-K1 cells stably transfected with hERG cDNA were obtained from GailRobertson at the University of Wisconsin. Cells were held in cryogenicstorage until needed. Cells were expanded and passaged in Dulbecco'sModified Eagles Medium/F12 supplemented with 10% fetal bovine serum and200 μg/1 mL geneticin. Cells were seeded onto poly-D-lysine (100 μg/mL)coated glass coverslips, in 35 mm² dishes (containing 2 mL medium) at adensity that enabled isolated cells to be selected for whole cellvoltage-clamp studies. The dishes were maintained in a humidified, 5%CO₂ environment at 37° C.

Extracellular solution was prepared at least every 7 days and stored at4° C. when not in use. The extracellular solution contained (mM): NaCl(137), KCl (4), CaCl₂ (1.8), MgCl₂ (1), Glucose (10),4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES) (10), pH 7.4with NaOH. The extracellular solution, in the absence or presence oftest compound, was contained in reservoirs, from which it flowed intothe recording chamber at approximately 0.5 mL/min. The intracellularsolution was prepared, aliquoted and stored at −20° C. until the day ofuse. The intracellular solution contained (mM): KCl (130), MgCl₂ (1),ethylene glycol-bis(beta-aminoethyl ether) N,N,N′,N′-tetra acetic acidsalt (EGTA) (5), MGATP (5),4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES) (10), pH 7.2with KOH. All experiments were performed at room temperature (20-22°C.).

The coverslips on which the cells were seeded were transferred to arecording chamber and perfused continuously. Gigaohm seals were formedbetween the cell and the patch electrode. Once a stable patch wasachieved, recording commenced in the voltage clamp mode, with theinitial holding potential at −80 mV. After a stable whole-cell currentwas achieved, the cells were exposed to test compound. The standardvoltage protocol was: step from the holding potential of −80 mV to +20mV for 4.8 sec, repolarize to −50 mV for 5 see and then return to theoriginal holding potential (−80 mV). This voltage protocol was run onceevery 15 sec (0.067 Hz). Peak current amplitudes during therepolarization phase were determined using pClamp software. Testcompounds at a concentration of 3 μM were perfused over the cells for 5minutes, followed by a 5-minute washout period in the absence ofcompound. Finally a positive control (cisapride, 20 nM) was added to theperfusate to test the function of the cell. The step from −80 mV to +20mV activates the hERG channel, resulting in an outward current. The stepback to −50 mV results in an outward tail current, as the channelrecovers from inactivation and deactivates.

Peak current amplitudes during the repolarization phase were determinedusing pCLAMP software. The control and test article data were exportedto Origin® (OriginLab Corp., Northampton Mass.) where the individualcurrent amplitudes were normalized to the initial current amplitude inthe absence of compound. The normalized current means and standarderrors for each condition were calculated and plotted versus the timecourse of the experiment.

Comparisons were made between the observed K⁺ current inhibitions afterthe five-minute exposure to either the test article or vehicle control(usually 0.3% DMSO). Statistical comparisons between experimental groupswere performed using a two-population, independent t-test (MicrocalOrigin v. 6.0). Differences were considered significant at p<0.05.

The smaller the percentage inhibition of the potassium ion current inthis assay, the smaller the potential for test compounds to change thepattern of cardiac repolarization when used as therapeutic agents. Thecompounds of the invention which were tested in this assay at aconcentration of 3 μM exhibited an inhibition of the potassium ioncurrent of less than about 20%, typically, less than about 15%.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto. Additionally, all publications, patents, andpatent documents cited hereinabove are incorporated by reference hereinin full, as though individually incorporated by reference.

1. A pharmaceutical composition comprising a pharmaceutically-acceptable carrier and a compound of formula (I):

or a pharmaceutically-acceptable salt or stereoisomer thereof, wherein: R¹ is hydrogen, halo, hydroxy, C₁₋₄alkyl, or C₁₋₄alkoxy; R² is C₃₋₄alkyl or C₃₋₆cycloalkyl; R³ is hydrogen or C₁₋₃alkyl; R⁴ is —S(O)₂—C₁₋₃alkyl, —C(O)O—C₁₋₃alkyl or —C(O)—C₁₋₃alkyl; n is an integer of 1, 2, or 3; Y is selected from: (a) a moiety of formula (a)

wherein: Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸, —N(R⁸)C(O)OR^(d), —N(R⁸)C(O)NR^(e)R^(f), —N(R⁸)SO₂NR^(g)R^(h), —C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —C(O)OR^(m), —OR⁸, —SR^(n), cyano, hydroxy-substituted C₁₋₄alkyl, hydroxy-substituted C₁₋₃alkoxy, —CF₃, pyridinyl, thiomorpholinyl, thiazolidinyl, imidazolyl, indolyl, tetrahydrofuranyl, pyrrolidinyl and piperidinyl, wherein pyrrolidinyl is optionally substituted with oxo and piperidinyl is optionally substituted with 1 to 3 halo; R⁵ is selected from hydrogen and C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, halo, and C₁₋₃alkoxy; m is an integer of 0, 1, 2, 3, 4, or 5; R⁶ and R⁷ are independently selected from hydrogen, hydroxy, halo and C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; R⁸ is independently hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; R^(a) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with —SO₂R^(b), C₃₋₆cycloalkyl or with from 1 to 3 halo; R^(c) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy and C₃₋₆cycloalkyl, or with from 1 to 3 halo; and R^(b), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i), R^(j), R^(k), R^(l), R^(m), and R^(n), are independently hydrogen or C₁₋₄alkyl; or R⁵ and R⁶, R⁵ and R⁸, or R⁶ and R⁸, taken together form a C₂₋₅alkylene, wherein the C₂₋₅alkylene is optionally substituted with 1 to 2 substituents selected from hydroxy, halo and C₁₋₄alkyl, wherein C₁₋₄ alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; or R⁸ and R^(a) taken together form a C₂₋₅alkylene; provided that when m is 1, Z forms a carbon-carbon bond with the carbon atom bearing the substituents R⁶ and R⁷; and when m is 0, Z is selected from —S(O₂)R⁸, —C(O)NR^(i)R^(j), —C(O)OR^(m), and —CF₃; and (c) —SR¹⁰, wherein R¹⁰ is hydrogen or C₁₋₄alkyl.
 2. The pharmaceutical composition of claim 1 wherein Y is selected from: (a) a moiety of formula (a), wherein Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸, —N(R⁸)C(O)NR^(e)R^(f), —C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —OR⁸, and cyano; and (c) —S—C₁₋₄alkyl.
 3. The pharmaceutical composition of claim 1 wherein Y is a moiety of formula (a).
 4. The pharmaceutical composition of claim 1 wherein the compound of formula (I) is a compound of formula (II):

or a pharmaceutically-acceptable salt or stereoisomer thereof, wherein: R¹ is hydrogen, halo, or C₁₋₃alkyl; R² is C₃₋₄alkyl; R³ is hydrogen or methyl, R⁴ is —S(O)₂—C₁₋₃alkyl, —C(O)O—C₁₋₃alkyl, or —C(O)—C₁₋₃alkyl; n is an integer of 1 or 2; R⁵ is selected from hydrogen and C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, halo, and C₁₋₃alkoxy; m is an integer of 1, 2, 3, 4, or 5; R⁶ and R⁷ are independently selected from hydrogen and C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸, —N(R⁸)C(O)OR^(d), —N(R⁸)C(O)NR^(e)R^(f), —N(R⁸)SO₂NR^(g)R^(h), —C(O)NR^(i)R^(j), —OC(O)NR^(k)R^(l), —C(O)OR^(m), —OR⁸, —SR^(n), and cyano; R⁸ is independently hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; R^(a) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with —SO₂R^(b), C₃₋₆cycloalkyl or with from 1 to 3 halo; R^(c) is hydrogen or C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy and C₃₋₆cycloalkyl, or with from 1 to 3 halo; and R^(b), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i), R^(j), R^(k), R^(l), R^(m), and R^(n), are independently hydrogen or C₁₋₄alkyl; or R⁵ and R⁶, R⁵ and R⁸, or R⁶ and R⁸, taken together form a C₂₋₅alkylene, wherein the C₂₋₅alkylene is optionally substituted with 1 to 2 substituents selected from hydroxy, halo and C₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with 1 to 2 substituents selected from hydroxy, C₁₋₃alkoxy, and cyano; or R⁸ and R^(a) taken together form a C₂₋₅alkylene.
 5. The pharmaceutical composition of claim 4 wherein R¹ is hydrogen, R² is C₃₋₄alkyl, and R³ is hydrogen.
 6. The pharmaceutical composition of claim 5 wherein R⁴ is selected from —S(O)₂CH₃, —C(O)OCH₃, and —C(O)CH₃.
 7. The pharmaceutical composition of claim 6 wherein n is 1, and m is 1, 2 or
 3. 8. The pharmaceutical composition of claim 7 wherein Z is selected from —N(R⁸)SO₂R^(a), —N(R⁸)C(O)R^(c), —S(O₂)R⁸, and —N(R⁸)C(O)NR^(e)R^(f).
 9. The pharmaceutical composition of claim 1 wherein the compound of formula (I) is selected from: 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid ((1S,3R,5R)-8-{3-[(2-ethylsulfanylethyl)methanesulfonylamino]-2-hydroxypropyl}-8-aza-bicyclo[3.2.1]oct-3-yl)amide; (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-{2-[3-(methanesulfonylmethylamino)piperidin-1-yl]ethyl}-carbamic acid methyl ester; [2-(3-acetylaminopiperidin-1-yl)ethyl]-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamic acid methyl ester; [2-(3-acetylaminopyrrolidin-1-yl)ethyl]-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamic acid methyl ester; (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(3-dimethylaminosulfonyl-aminopyrrolidin-1-yl)-ethyl]-carbamic acid methyl ester; (2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)-amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-[2-(3-dimethylaminosulfonyl-methylaminopyrrolidin-1-yl)-ethyl]-carbamic acid methyl ester; {2-[3-(1,1-dioxo-1λ⁶-isothiazolidin-2-yl)pyrrolidin-1-yl]ethyl}-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-aza-bicyclo[3.2.1]oct-8-yl}propyl)-carbamic acid methyl ester; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid [(1S,3R,5R)-8-(3-{acetyl-[2-(3-carbamoylpiperidin-1-yl)ethyl]amino}-2-hydroxypropyl)-8-azabicyclo[3.2.1]oct-3-yl]-amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-{2-[4-(methanesulfonylaminomethyl)piperidin-1-yl]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide; (1-{2-[acetyl-(2-hydroxy-3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)amino]-ethyl}pyrrolidin-3-yl)-carbamic acid methyl ester; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-{2-[3-(dimethylaminosulfonylmethylamino)pyrrolidin-1-yl]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-{2-[3-(trimethylureido)pyrrolidin-1-yl]ethyl}amino)-2-hydroxy-propyl]-8-azabicyclo-[3.2.1]oct-3-yl}amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-{2-[(1-dimethylsulfamoylpyrrolidin-3-yl)methylamino]ethyl}amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid ((1S,3R,5R)-8-{3[acetyl-(2-{[2-(methanesulfonylmethylamino)ethyl]methylamino}-ethyl)amino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-{2-[(2-methanesulfonylaminoethyl)methylamino]ethyl}amino)-2-hydroxypropyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide; and 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid ((1S,3R,5R)-8-{3-[acetyl-(2-{[2-(1,1-dioxo-1λ⁶-isothiazolidin-2-yl)ethyl]methylamino}ethyl)amino]-2-hydroxypropyl}-8-azabicyclo[3.2.1]oct-3-yl)amide; and pharmaceutically-acceptable salts and stereoisomers thereof.
 10. A method of treating a mammal having a disease or medical condition mediated by treatment with a 5-HT₄ receptor agonist, wherein the disease or medical condition is chronic constipation, constipation-predominant irritable bowel syndrome, functional dyspepsia, gastroparesis, gastroesophageal reflux disease, or drug-induced delayed transit, the method comprising administering to the mammal, the pharmaceutical composition of claim
 1. 11. The method of claim 10 wherein the disease or medical condition is chronic constipation or constipation-predominant irritable bowel syndrome.
 12. The method of claim 10 wherein the disease or medical condition is functional dyspepsia, gastroparesis, gastroesophageal reflux disease, or drug-induced delayed transit.
 13. A method of treating a disorder of reduced motility of the gastrointestinal tract in a mammal, the method comprising administering to the mammal, the pharmaceutical composition of claim
 1. 14. A method of treating a mammal having a disease or medical condition mediated by treatment with a 5-HT₄ receptor agonist, wherein the disease or medical condition is chronic constipation, constipation-predominant irritable bowel syndrome, functional dyspepsia, gastroparesis, gastroesophageal reflux disease, or drug-induced delayed transit, the method comprising administering to the mammal, the pharmaceutical composition of claim
 4. 15. The method of claim 14 wherein the disease or medical condition is chronic constipation or constipation-predominant irritable bowel syndrome.
 16. The method of claim 14 wherein the disease or medical condition is functional dyspepsia, gastroparesis, gastroesophageal reflux disease, or drug-induced delayed transit. 